Nothing Special   »   [go: up one dir, main page]

EP0247091B1 - Modulare einheit von antikörpergenen, daraus hergestellte antikörper und verwendung - Google Patents

Modulare einheit von antikörpergenen, daraus hergestellte antikörper und verwendung Download PDF

Info

Publication number
EP0247091B1
EP0247091B1 EP86906676A EP86906676A EP0247091B1 EP 0247091 B1 EP0247091 B1 EP 0247091B1 EP 86906676 A EP86906676 A EP 86906676A EP 86906676 A EP86906676 A EP 86906676A EP 0247091 B1 EP0247091 B1 EP 0247091B1
Authority
EP
European Patent Office
Prior art keywords
region
sequence
human
mouse
cdna
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Revoked
Application number
EP86906676A
Other languages
English (en)
French (fr)
Other versions
EP0247091A1 (de
EP0247091A4 (de
Inventor
Randy R. Robinson
Alvin Y. Liu
Arnold H. Horwitz
Randolph Wall
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Xoma Corp
Original Assignee
Xoma Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=25161322&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=EP0247091(B1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Xoma Corp filed Critical Xoma Corp
Priority to AT86906676T priority Critical patent/ATE95235T1/de
Publication of EP0247091A4 publication Critical patent/EP0247091A4/de
Publication of EP0247091A1 publication Critical patent/EP0247091A1/de
Application granted granted Critical
Publication of EP0247091B1 publication Critical patent/EP0247091B1/de
Anticipated expiration legal-status Critical
Revoked legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3046—Stomach, Intestines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C07K16/461—Igs containing Ig-regions, -domains or -residues form different species
    • C07K16/462—Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
    • C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/88—Lyases (4.)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence

Definitions

  • This invention relates to recombinant DNA methods of preparing immunoglobulins, genetic sequences coding therefor, as well as methods of obtaining such sequences.
  • the present invention bridges both the hybridoma and monoclonal antibody technologies and provides a quick and efficient method, as well as products derived therefrom, for the improved production of chimeric human/non-human antibodies, or of "class switched" antibodies.
  • Cabilly et al. disclose in EP-A-125023, the construction of an expression vector for a chimeric immunoglobulin chain which comprises repeated steps of digestion with restriction endonuclease, purification of fragments or filling-in with Klenow-polymerase and dNTP's and ligation.
  • the resulting plasmid carries a human/mouse hybrid gene, comprising a mouse variable region and a human constant region.
  • GB-A-2137631 discloses cDNA clones encoding immunoglobulin lambda light chains and immunoglobulin j1. heavy chains.
  • EP-A-184187 (prior art under Article 54 (3) EPC) the construction of mouse- human chimeric immunoglobulin heavy chain DNA, which DNA is isolated from genomic DNAs of mouse and human, respectively.
  • Boulianne G. L. et al., Nature, 312: 643 (December 13, 1984), also produced antibodies consisting of mouse variable regions joined to human constant regions. They constructed immunoglobulin genes in which the DNA segments encoding mouse variable regions specific for the hapten trinitrophenyl (TNP) were joined to genomic DNA segments encoding human mu and kappa constant regions. These chimeric genes were expressed as functional TNP binding chimeric IgM.
  • TNP hapten trinitrophenyl
  • NP 4-hydroxy-3-nitrophenacetyl
  • Taniguchi M., in European Patent Publication No. 171 496 (published February 19, 1985) discloses the production of chimeric antibodies having variable regions with tumor specificity derived from experimental animals, and constant regions derived from human.
  • the corresponding heavy and light chain genes are produced in the genomic form, and expressed in mammalian cells.
  • Boss, M. A., European Patent Application 120694 shows expression in E. coli of non-chimeric immunoglobulin chains with 4-nitrophenyl specificity. There is a broad description of chimeric antibodies but no details (see p. 9).
  • Seno M. et al., Nucleic Acids Research, 11: 719-726 (1983), describe the preparation of a cDNA sequence and recombinant plasmids containing the same coding for part of the variable region and all of the constant region of the human IgE heavy chain (epsilon chain).
  • the invention provides a novel approach for producing genetically engineered antibodies of desired variable region specificity and constant region properties through gene cloning and expression of light and heavy chains.
  • the cloned immunoglobulin gene products can be produced by expression in genetically engineered organisms.
  • the application of chemical gene synthesis, recombinant DNA cloning, and production of specific immunoglobulin chains in various organisms provides an effective solution for the efficient large scale production of human monoclonal antibodies with the antigen specificities of either human or non-human, especially rodent, monoclonal antibodies.
  • the invention also provides a solution to the problem of class switching antibody molecules, so as to readily prepare immunoglobulins of a certain binding specificity of any given class.
  • the invention provides vectors comprising cDNA sequences coding for immunoglobulin chains comprising a constant human region and a variable, either human or non-human, region.
  • the immunoglobulin chains can either be heavy or light.
  • the invention also provides gene sequences coding for immunoglobulin chains comprising a cDNA variable region of non-human origin and a genomic constant region of human origin.
  • the invention also provides sequences as above, present in recombinant DNA molecules, especially in vehicles such as plasmid vectors, capable of expression in desired prokaryotic or eukaryotic hosts.
  • the invention also provides consensus sequences and specific oligonucleotide sequences useful as probes for hybridization and priming cDNA synthesis of any hybridoma mRNA coding for variable regions of any desired specificity.
  • the invention provides hosts capable of producing, by culture, chimeric antibodies and methods of using these hosts.
  • the invention also provides chimeric immunoglobulin individual chains and whole assembled molecules having human constant regions and non-human variable regions, wherein both variable regions have the same binding specificity.
  • immunoglobulin chains and/or molecules provided by the invention are:
  • the resulting antibody molecule binds to two different antigens.
  • the invention also provides for a genetic sequence, especially a cDNA sequence, coding for the variable region of an antibody molecule heavy and/or light chain, operably linked to a sequence coding for a polypeptide different than an immunoglobulin chain (e.g., an enzyme).
  • a genetic sequence especially a cDNA sequence, coding for the variable region of an antibody molecule heavy and/or light chain, operably linked to a sequence coding for a polypeptide different than an immunoglobulin chain (e.g., an enzyme).
  • cDNA sequences are particularly advantageous over genomic sequences (which contain introns), in that cDNA sequences can be expressed in bacteria or other hosts which lack RNA splicing systems.
  • preferred specific antibodies are those having specificities to cancer-related antigens.
  • antibodies are composed of two light and two heavy chain molecules.
  • Light and heavy chains are divided into domains of structural and functional homology.
  • the variable regions of both light (V L ) and heavy (V H ) chains determine recognition and specificity.
  • the constant region domains of light (C L ) and heavy (C H ) chains confer important biological properties such as antibody chain association, secretion, transplacental mobility, complement binding, and the like.
  • V H , D and J H ; or V L and J L germ line gene segments
  • V H , D and J H ; or V L and J L germ line gene segments
  • V L -J L and V H -D-J H V segments are joined by DNA rearrangements to form the complete V regions expressed in heavy and light chains respectively.
  • the rearranged, joined (V L -J L and V H -D-J H ) V segments then encode the complete variable regions or antigen binding domains of light and heavy chains, respectively.
  • the invention provides a novel approach for the cloning and production of human antibodies with desired specificity. Generally, the method combines five elements:
  • the invention employs cloned hybridoma B cell lines producing monoclonal antibodies of defined specificity for the isolation of mRNA for cDNA cloning. Because many lymphoid cell lines contain highly active nucleases which degrade mRNA during isolation, the invention uses mRNA preparation methods specifically developed for the isolation of intact mRNA from cells and tissues containing active nucleases. One such method yields total RNA preparations by cell or tissue disruption in an ethanol-perchlorate dry ice mixture which reduces nuclease action (Lizardi, P. M. et al., Anal. Biochem., 98: 116 (1979)). This method gives intact translatable mRNA.
  • RNA extraction methods include extraction of cells with lithium chloride plus urea (Auffray, C., and Rougeon, F., Eur. J. Biochem., 107: 303 (1980)) or guanidine thiocyanate (Chirgwin, J. M. et al., Biochemistry, 18: 5294 (1979)) to prepare total RNA.
  • J region i.e. joining region, see Figure 1.
  • Heavy and light chain J regions have different sequences, but a high degree of sequence homology exists (greater than 80%) within the heavy J H regions or the kappa light chain J regions.
  • the invention provides consensus sequences of light and heavy chain J regions useful in the design of oligonucleotides (designated herein as UlGs) for use as primers or probes for cloning immunoglobulin light or heavy chain mRNAs or genes ( Figures 2 or 7).
  • UIG may be capable of hybridizing to all immunoglobulin mRNAs or genes containing a single specific J sequence, such as UIG-MJH3 which detects only mouse J H 3 sequences ( Figure 7).
  • UIG design can also include a sequence to introduce a restriction enzyme site into the cDNA copy of an immunoglobulin gene (see Figure 7).
  • the invention may, for example, utilize chemical gene synthesis to generate the UIG probes for the cloning of V regions in immunoglobulin mRNA from hybridoma cells making monoclonal antibodies of desired antigen specificities.
  • a multi-stage procedure is utilized for generating complete V + C region cDNA clones from hybridoma cell light and heavy chain mRNAs.
  • the invention utilizes UIG probes as "primers" for reverse transcriptase copying of the complete V region and leader coding sequences of heavy and light chain mRNAs ( Figure 3).
  • the complementary strand of the primer extended cDNA is then synthesized, and this double-stranded cDNA is cloned in appropriate cDNA cloning vectors such as pBR322 (Gubler and Hoffman, Gene, 25: 263 (1983)) or pQ23 ( Figure 5; Maniatis, T.
  • Clones are screened for specific hybridization with UIG oligonucleotide probes. Positive heavy and light chain clones identified by this screening procedure are mapped and sequenced to select those containing V region and leader coding sequences.
  • An alternative method is to make cDNA clones using oligo-dT as a primer, followed by selection of light and heavy chain clones by standard hybridization methods.
  • a second stage utilizes cloning of C region gene segments to form heavy and light chain module vectors.
  • cDNA clones of human heavy and light chain immunoglobulin mRNA are prepared. These cDNA clones are then converted into C region module vectors by site-directed mutagenesis to place a restriction site at a desired location near a boundary of the constant region.
  • An alternative method utilizes genomic C region clones as the source for C region module vectors.
  • a third stage of cDNA cloning involves the generation of complete light and heavy chain coding sequences with linked V and C regions.
  • the cloned V region segments generated as above are excised and ligated to light or heavy chain C region module vectors.
  • Operationally linked means in-frame joining of coding sequences to derive a continuously translatable gene sequence without alterations or interruptions of the triplet reading frame.
  • cDNA genetic sequences in the present invention are the fact that they code continuously for immunoglobulin chains, either heavy or light.
  • continuous is meant that the sequences do not contain introns (i.e. are not genomic sequences, but rather, since derived from mRNA by reverse transcription, are sequences of contiguous exons).
  • This characteristic of the cDNA sequences provided by the invention allows them to be expressible in prokaryotic hosts, such as bacteria, or in lower eukaryotic hosts, such as yeast.
  • Another advantage of cDNA cloning methods is the ease and simplicity of obtaining V region gene modules.
  • non-human as used in the invention is meant to include any animal other than a human, wherein an immune response can be generated which then leads to usable B cells resulting in corresponding hybridomas or B cell clones obtained by viral transformation and the like.
  • Such animals commonly include rodents such as the mouse or the rat. Because of ease of preparation and great availability, the mouse is at present the preferred, non-human animal.
  • Mouse-mouse hybridomas are thus utilized as the preferred sources for heavy and light chain variable regions.
  • the invention provides entire V and/or C region cDNA sequences. This means that the sequences code for substantially operable V and/or C regions, without lacking any major structural portions thereof.
  • variable and constant are used functionally to denote those regions of the immunoglobulin chain, either heavy or light chain, which code for properties and features possessed by the variable and constant regions in natural non-chimeric antibodies. As noted, it is not necessary for the complete coding region for variable or constant regions to be present, as long as a functionally operating region is present and available.
  • hybridomas secreting antibodies which are reactive with viral antigens, including Dengue complex specific (ATCC HB 114), Dengue type 1 virus (ATCC HB 47), Dengue type 2 virus (ATCC HB 46), Dengue type 3 virus (ATCC HB 49), Dengue type 4 virus (ATCC HB 48), Epstein-Barr receptor (ATCC HB 135), Flavivirus group (ATCC HB 112), hepatitis B surface antigen (ATCC CRL 8017 and 8018), herpes simplex type I (ATCC HB 8068), herpes simplex type II (ATCC HB 8067), influenza virus (ATCC CL 189), influenza A virus, matrix protein (ATCC HB 64), influenza A virus, nucleoprotein (ATCC HB 65), influenza A Bangkok/1/79HA (ATCC HB 66), influenza AWSN NP (ATCC HB 67), SV40 large T antigen (ATCC TIB 115), SV40 large T antigen, C-terminal end (ATCC HB 114
  • hybridomas examples include those secreting antibodies to tumor associated antigens or to human lymphocyte antigens, such as those reactive to human tumor-associated CEA, high mw (ATCC CRL 8019); human tumor-associated alpha-fetoprotein, IgG 1 K - (ATCC HB 134); human B lymphocyte HLA-DR, monomorphic, IgG 2b (ATCC HB 104); human T lymphocyte T cell precursors, IgG 1 (ATCC CRL 8022); human T lymphocyte T cell subset, helper, IgG 2b (ATCC CRL 8002); T subset, suppressor/cytotoxic, human, IgG 1 (ATCC CRL 8013); T cell subset, suppressor/cytotoxic, human, IgG 2a (ATCC CRL 8014); T cells, peripheral, human, IgG 1 (ATCC CRL 8000); T cells, peripheral, human, IgG 2a (ATCC CRL 8001); thymocytes, "common,” human, IgG 1
  • Expression vehicles include plasmids or other vectors. Preferred among these are vehicles carrying a functionally complete human constant heavy or light chain sequence having appropriate restriction sites engineered so that any variable heavy or light chain sequence with the appropriate cohesive ends can be easily inserted thereinto. Human constant heavy or light chain sequence-containing vehicles are thus an important embodiment of the invention. These vehicles can be used as intermediates for the expression of any desired complete heavy or light chain in any appropriate host.
  • yeast provides substantial advantages for the production of immunoglobulin light and heavy chains. Yeasts carry out post-translational peptide modifications including glycosylation. A number of recombinant DNA strategies now exist which utilize strong promoter sequences and high copy number plasmids which can be used for overt production of the desired proteins in yeast. Yeast recognizes leader sequences on cloned mammalian gene products and secretes peptides bearing leader sequences (i.e. prepeptides) (Hitzman, et al., 11th International Conference on Yeast, Genetics and Molecular Biology, Montpelier, France, September 13-17, 1982).
  • Yeast gene expression systems can be routinely evaluated for the level of heavy and light chain production, protein stability, and secretion. Any of a series of yeast gene expression systems incorporating promoter and termination elements from the actively expressed genes coding for glycolytic enzymes produced in large quantities when yeasts are grown in mediums rich in glucose can be utilized. Known glycolytic genes can also provide very efficient transcription control signals. For example, the promoter and terminator signals of the iso-1-cytochrome C (CYC-1) gene can be utilized.
  • CYC-1-cytochrome C CYC-1-cytochrome C
  • E. coli K12 strain 294 (ATCC 31446) is particularly useful.
  • Other microbial strains which may be used include E. coli X1776 (ATCC 31537).
  • the aforementioned strains, as well as E. coli W3110 (ATCC 27325) and other enterobacteria such as Salmonella typhimurium or Serratia marcescens, and various Pseudomonas species may be used.
  • plasmid vectors containing replicon and control sequences which are derived from species compatible with a host cell are used in connection with these hosts.
  • the vector ordinarily carries a replication site, as well as specific genes which are capable of providing phenotypic selection in transformed cells.
  • E. coli is readily transformed using pBR322, a plasmid derived from an E. coli species (Bolivar, et al., Gene, 2: 95 (1977)).
  • pBR322 contains genes for ampicillin and tetracycline resistance, and thus provides easy means for identifying transformed cells.
  • the pBR322 plasmid or other microbial plasmids must also contain, or be modified to contain, promoters which can be used by the microbial organism for expression of its own proteins.
  • promoters most commonly used in recombinant DNA construction include the beta-lactamase (penicillinase) and lactose (beta-galactosidase) promoter systems (Chang et al., Nature, 275: 615 (1978); Itakura et al., Science, 198:1056 (1977)); and tryptophan promoter systems (Goeddel et al., Nucleic Acids Research, 8: 4057 (1980); EPO Publication No. 0036776). While these are the most commonly used, other microbial promoters have been discovered and utilized.
  • a genetic construct for any heavy or light chimeric immunoglobulin chain can be placed under the control of the leftward promoter of bacteriophage lambda (P L ).
  • This promoter is one of the strongest known promoters which can be controlled. Control is exerted by the lambda repressor, and adjacent restriction sites are known.
  • lactose dependent E. coli chromosomal DNA comprises a lactose or lac operon which mediates lactose digestion by elaborating the enzyme beta-galactosidase.
  • the lac control elements may be obtained from bacteriophage lambda pLAC5, which is infective for E. coli.
  • the lac promoter-operator system can be induced by IPTG.
  • promoter/operator system or portions thereof can be employed as well.
  • arabinose, colicine E1, galactose, alkaline phosphatase, tryptophan, xylose, tac, and the like can be used.
  • Mammalian cells are mammalian cells, grown in vitro in tissue culture, or in vivo in animals. Mammalian cells provide post-translational modifications to immunoglobulin protein molecules including leader peptide removal, correct folding and assembly of heavy and light chains, glycosylation at correct sites, and secretion of functional antibody protein from the cell as H 2 L 2 molecules.
  • Mammalian cells which may be useful as hosts for the production of antibody proteins include cells of fibroblast origin, such as Vero (ATCC CRL 81) or CHO-K1 (ATCC CRL 61), or cells of lymphoid origin, such as the hybridoma Sp2/0-Ag14 (ATCC CRL 1581) or the myeloma P3X63Ag8 (ATCC TIB 9), and its derivatives.
  • fibroblast origin such as Vero (ATCC CRL 81) or CHO-K1 (ATCC CRL 61)
  • cells of lymphoid origin such as the hybridoma Sp2/0-Ag14 (ATCC CRL 1581) or the myeloma P3X63Ag8 (ATCC TIB 9), and its derivatives.
  • vectors utilize DNA elements which provide an autonomously replicating extrachromosomal plasmid, derived from animal viruses, such as bovine papillomavirus (Sarver, N. et al., Proc. Natl. Acad. Sci., USA, 79: 7147 (1982)), polyoma virus (Deans, R. J. et al., Proc. Natl. Acad. Sci., USA, 81: 1292 (1984)), or SV40 virus (Lusky, M. and Botchan, M., Nature, 293: 79 (1981)).
  • bovine papillomavirus Sesarver, N. et al., Proc. Natl. Acad. Sci., USA, 79: 7147 (1982)
  • polyoma virus Deans, R. J. et al., Proc. Natl. Acad. Sci., USA, 81: 1292 (1984)
  • SV40 virus Lusky, M. and Botchan
  • a second class of vectors relies upon the integration of the desired gene sequences into the host cell chromosome.
  • Cells which have stably integrated the introduced DNA into their chromosomes can be selected by also introducing drug resistance genes such as E. coli gpt (Mulligan, R. C. and Berg, P., Proc. Natl. Acad. Sci., USA, 78: 2072 (1981)) or Tn5 neo (Southern, P. J. and Berg, P., J. Mol. Appl. Genet., 1: 327 (1982)).
  • the selectable marker gene can be either directly linked to the DNA gene sequences to be expressed, or introduced into the same cell by co-transfection (Wigler, M. et al., Cell, 16: 77 (1979)).
  • an immunoglobulin cDNA is comprised only of sequences representing the mature mRNA encoding an antibody protein or its precursor, additional gene expression elements regulating transcription of the gene and processing of the RNA are required for optimal synthesis of immunoglobulin mRNA.
  • These elements may include splice signals, as well as transcription promoters including inducible promoters, enhancers, and termination signals.
  • cDNA expression vectors incorporating such elements include those described by Okayama, H. and Berg, P., Mol. Cell Biol., 3: 280 (1983); Cepko, C. L. et al., Cell, 37: 1053 (1984); and Kaufman, R. J., Proc. Natl. Acad. Sci., U g A - ,-82. - 689 (1985).
  • An approach to evaluate optimal vectors for the expression of immunoglobulin cDNA in mammalian cells involves first placing the immunoglobulin DNA sequences into vectors capable of stably integrating into the cell genome, or replicating autonomously as an extrachromosomal plasmid.
  • the vectors can be used to evaluate different gene expression elements for optimal immunoglobulin synthesis.
  • mammalian cells may express chimeric immunoglobulin genes which are comprised of a variable region cDNA module plus a constant region which is composed in whole or in part of genomic sequences.
  • a variable region cDNA module plus a constant region which is composed in whole or in part of genomic sequences.
  • human constant region genomic clones have been described (Ellison, J. W. et al., Nucl. Acids Res., 10: 4071 (1982), or Max, E. et al., Cell, 29: 691 (1982)).
  • the use of such genomic sequences may be convenient for the simultaneous introduction of immunoglobulin enhancers, splice signals, and transcription termination signals along with the constant region gene segment.
  • human IgM antibodies can be transmuted to human IgG antibodies by preparing constructs containing human constant IgG cDNA or genomic sequences, linked to variable human cDNA sequences obtained from a cell producing the original IgM antibody. These constructs are then introduced into appropriate hosts and expressed.
  • the invention provides "chimeric" immunoglobulin chains, either heavy or light.
  • a chimeric chain contains a constant region substantially similar to that present in the heavy chain of a natural human immunoglobulin, and a variable region having any desired antigenic specificity.
  • the variable region is either from human or non-human origin.
  • the invention also provides immunoglobulin molecules having heavy and light chains associated so that the overall molecule exhibits desired binding and recognition properties.
  • immunoglobulin molecules having heavy and light chains associated so that the overall molecule exhibits desired binding and recognition properties.
  • Various types of immunoglobulin molecules are provided: monovalent, divalent, dispecific (i.e., with different variable regions), molecules with chimeric heavy chains and non-chimeric light chains, or molecules with variable binding domains attached to peptide moieties carrying desired functions.
  • Antibodies having chimeric heavy chains of the same or different variable region binding specificity and non-chimeric (i.e., all human or all non-human) light chains can be prepared by appropriate association of the needed polypeptide chains. These chains are individually prepared by the modular assembly methods of the invention.
  • the antibodies of the invention having human constant region can be utilized for passive immunization, especially in humans, without negative immune reactions such as serum sickness or anaphylactic shock.
  • the antibodies can, of course, also be utilized in prior art immunodiagnostic assays and kits, in labelled form for in vivo imaging, wherein the label can be a radioactive emitter, or an NMR contrasting agent such as a carbon-13 nucleus, or an X-ray contrasting agent, such as a heavy metal nucleus.
  • the antibodies can also be used for in vitro localization of antigens by appropriate labelling.
  • the antibodies can be used for therapeutic purposes by themselves in complement mediated lysis or can be coupled to toxins or toher therapeutic moieties.
  • Class switching of antibodies is useful when it is desired to change the association, aggregation or other properties of antibodies obtained from cell fusion or hybridoma technology.
  • most human- human monoclonals are of the IgM class, which are known for their ease of reduction and aggregation. Changing such antibodies to other antibody types, such as IgG, IgA or IgE, is thus of great benefit.
  • Mixed antibody-enzyme molecules can be used for immunodiagnostic methods, such as ELISA.
  • Mixed antibody-peptide effector conjugates can be used for targeted delivery of the effector moiety with a high degree of efficacy and specificity.
  • the human cell lines GM2146 and GM1500 were obtained from the Human Mutant Cell Repository (Camden, New Jersey) and cultured in RPM11640 plus 10% fetal bovine serum (M. A. Bioproducts).
  • the cell lines Sp2/0 and CRL 8017 were obtained from the American Type Culture Collection and grown in Dulbecco's Modified Eagle Medium (DMEM) plus 4.5 g/I glucose (M. A. Bioproducts) plus 10% fetal bovine serum (Hyclone, Sterile Systems, Logan, Utah). Media were supplemented with penicillin/streptomycin (Irvine Scientific, Irvine, California).
  • the plasmids pBR322, pL1 and pUC12 were purchased from Pharmacia P-L Biochemicals (Milwaukee, Wisconsin).
  • the plasmids pSV2-neo and pSV2-gpt were obtained from BRL (Gaithersburg, Maryland), and are available from the American Type Culture Collection (Rockville, Maryland).
  • pHu-gamma-I is a subclone of the 8.3 Kb Hindlll to BamHl fragment of the human IgGI chromosomal gene. A separate isolation of the human IgGI chromosomal gene is described by Ellison, J. W. et al., Nucl. Acids Res., 10: 4071 (1982).
  • M8alphaRX12 contains the 0.7 Kb Xbal to EcoRl fragment containing the mouse heavy chain enhancer from the J-C intron region of the M603 chromosomal gene (Davis, M. et al., Nature, 283: 733) inserted into M13mp10.
  • G-tailed pUC9 was purchased from Pharmacia P-L.
  • DNA manipulations involving purification of plasmid DNA by buoyant density centrifugation, restriction endonuclease digestion, purification of DNA fragments by agarose gel electrophoresis, ligation and transformation of E. coli were as described by Maniatis, T. et al., Molecular Cloning: A Laboratory Manual, (1982). Restriction endonucleases and other DNA/RNA modifying enzymes were purchased from Boehringer-Mannheim (Indianapolis, Indiana), BRL, New England Biolabs (Beverly, Massachusetts) and Pharmacia P-L.
  • Oligonucleotides were either synthesized by the triester method of Ito et al. (Nucl. Acids Res., 10: 1755 (1982)), or were purchased from ELESEN, Los Angeles, California. Tritylated, deblocked oligonucleotides were purified on Sephadex-G50, followed by reverse-phase HPLC with a 0-25% gradient of acetonitrile in 10mM triethylamine-acetic acid, pH 7.2, on a C18 uBondapak column (Waters Associates). Detritylation was in 80% acetic acid for 30 min., followed by evaporation thrice. Oligonucleotides were labeled with [gamma- 32 P]ATP plus T4 polynucleotide kinase.
  • Total cellular RNA was prepared from tissue culture cells by the method of Auffray, C. and Rougeon, F. (Eur. J. Biochem., 107: 303 (1980)) or Chirgwin, J. M. et al. (Biochemistry, 18: 5294 (1979)). Preparation of poly(A) + RNA, methyl-mercury agarose gel electrophoresis, and "Northern" transfer to nitrocellulose were as described by Maniatis, T. et al., supra. Total cellular RNA or poly(A) + RNA was directly bound to nitrocellulose by first treating the RNA with formaldehyde (White, B. A. and Bancroft, F. C., J. Biol.
  • Hybridization to filterbound RNA was with nick-translated DNA fragments using conditions described by Margulies, D. H. et al. (Nature, 295: 168 (1982)) or with 32 P-labelled oligonucleotide using 4xSSC, 10X Denhardt's, 100 ug/ml salmon sperm DNA at 37 °C overnight, followed by washing in 4xSSC at 37 ° C.
  • Oligo-dT primed cDNA libraries were prepared from poly(A) + RNA from GM1500 and GM2146 cells by the methods of Land, H. et al. (Nucl. Acids Res., 9: 2251 (1981)) and Gubler, V. and Hoffman, B. J., Gene, 25: 263 (1983), respectively.
  • the cDNA libraries were screened by in situ hybridization (Maniatis, T., supra) with 32 P-labelled oligonucleotides using the conditions shown above, or with nick-translated DNA fragments using the conditions of de Lange et al. (Cell, 34: 891 (1983)).
  • Poly(A) + RNA (20 ug) was mixed with 1.2 ug primer in 40 ul of 64mM KCI. After denaturation at 90°C for 5 min. and then chilling in ice, 3 units Human Placental Ribonuclease Inhibitor (BRL) was added in 3 ul of 1 M Tris-HCI, pH 8.3. The oligonucleotide was annealed to the RNA at 42°C for 15 minutes, then 12 ul of .05M DTT, .05M MgC1 2 , and 1 mM each of dATP, dTTP, dCTP, and dGTP was added. 2 ul of alpha- 32 p-dATp (400 Ci/mmol, New England Nuclear) was added, followed by 3 ul of AMV reverse transcriptase (19 units/ul, Life Sciences).
  • BRL Human Placental Ribonuclease Inhibitor
  • RNA-DNA hybrid was extracted with phenol, then with chloroform, and precipitated with ethanol.
  • Second strand synthesis, homopolymer tailing with dGTP or dCTP, and insertion into homopolymer tailed vectors was as described by Gubler and Hoffman, supra.
  • Single stranded M13 subclone DNA (1 ug) was combined with 20 ng oligonucleotide primer in 12.5 ul of Hin buffer (7 mM Tris-HCI, pH 7.6, 7 mM MgCI 2 , 50 mM NaCI). After heating to 95 ° C in a sealed tube, the primer was annealed to the template by slowly cooling from 70° C to 37° C for 90 minutes. 2 ul dNTPs (1 mM each), 1 ul 32 P-dATP (10 uCi), 1 ul DTT (0.1 M) and 0.4 ul Klenow DNA Poll (2u, Boehringer Mannheim) were added and chains extended at 37° C for 30 minutes.
  • Bacterial protoplasts (10 ml) were mixed with 10x10 6 Sp2/0 cells and pelleted by centrifugation at 4,000 x g at 22° C for 20 min. After pipetting off the supernatant, the pellet was suspended in the remaining drop of medium by flicking the tube. 2ml of 10% DMSO, 37% (w/v) PEG6000 (Kodak) in DMEM was added dropwise with mixing over 45 sec. After 15 sec., 2 ml of 42% PEG6000 in DMEM was added over 45 sec. Complete DMEM (45 ml) was slowly added with mixing. Cells were pelleted at 2500 x g, then washed and pelleted thrice.
  • Cytoplasmic protein extract was prepared by vortexing 1x10 6 cells in 160 ul of 1% NP40@, 0.15 M NaCI, 10 mM Tris, 1 mM EDTA, pH 7.6 at 0 ° C, 15 minutes, followed by centrifugation at 10,000 x g to remove insoluble debris.
  • Double antibody sandwich ELISA (Voller, A. et al., in Manual of Clinical Immunology, 2nd Ed., Eds. Rose, N. and Friedman, H., pp. 359-371, 1980) using affinity purified antisera was used to detect specific immunoglobulins.
  • the plate-bound antiserum is goat anti-human IgG (KPL, Gaithersburg, Maryland) at 1/1000 dilution, while the peroxidase-bound antiserum is goat anti-human IgG (KPL or Tago, Burlingame) at 1/4000 dilution.
  • the plate-bound antiserum is goat anti-human kappa (Tago) at 1/500 dilution, while the peroxidase-bound antiserum is goat anti-human kappa (Cappel) at 1/1000 dilution.
  • Antibodies binding hepatitis B surface antigen were detected using a commercial (Abbott, AUSAB) assay.
  • the following examples show the preparation of chimeric antibodies each having a human constant region and a non-human variable region. These examples outline the step-by-step process of preparing the chimeric antibodies.
  • the cell line GM2146 was used as the source in mRNA preparation and cDNA cloning. This cell line secretes IgGI (Simmons, J. G. et al., Scand. J. Immunol., 14: 1-13, 1981). Tests of this cell line indicated that it secretes IgA as well as IgG.
  • the cell line was cloned, and results indicated that five of six subclones secreted IgG only, while one of six subclones secreted IgA only.
  • Poly(A) + RNA was prepared from the cell line and a cDNA library was prepared from the poly(A) + RNA by the method of Gubler, U. and Hoffman, B. J., Gene, 25: 263-269 (1983).
  • An initial plating of the cDNA transformed into E. coli strains HB101 and RR1 yielded a total of 1500 colonies, which were screened by hybridization to a Hindlll to BamHl fragment of a genomic clone of human IgGI (pHu-gamma-1 Four positive clones were found.
  • Clone pGMH-6 provided the IgGI constant region module in the construction of cloning vectors for heavy chain variable region cloning.
  • a human cell line (GM1500) producing IgG 2 K was selected for the initial cloning phase.
  • Poly(A) + RNA prepared from GM1500 is active in in vitro translation using rabbit reticulocyte extracts.
  • a cDNA library was prepared from this RNA by the method of Land et al., Nucl. Acids Res., 9: 2251-2266 (1981), utilizing Kpnl digested and dG-tailed pQ23 as the cloning vector ( Figure 5). This vector contains Bglll, Kpnl and Sstl sites inserted between the BamHl and Sall sites of pBR322.
  • UIG-HuK a DNA probe, UIG-HuK, was synthesized and purified.
  • the UIG-HuK oligonucleotide has the sequence 5'-AGCCACAGTTCGTTT-3', and is designed to hybridize to all functional human kappa mRNA species at the J-C junction. This probe was used to prime cDNA synthesis on GM1500 RNA in the presence of dideoxynucleotides and reverse transcriptase.
  • cDNA clones positive for hybridization to the light chain probe were selected. Since the probe hybridizes to the J-C junction, the most important point was to determine if the clones had complete constant region sequence in addition to the J region.
  • Insert sizes for the two largest kappa cDNA clones were 0.6 and 0.9 kb; restriction enzyme mapping indicated that the entire constant region coding sequence was present in both clones ( Figure 6).
  • the human kappa cDNA clone pK2-3 was used to make the light chain constant region vector plNG2001 by inserting the Sau3A fragment comprising the human kappa constant and J regions into the Bcll site of pBR325 ( Figure 6B).
  • a variant of the human kappa cDNA clone was made by placing a Hindlll site in the J region. This was carried out by in vitro mutagenesis using a J K HINDIII oligonucleotide primer ( Figure 7c). The resultant plasmid is pGML60.
  • This vector was constructed from pUC12 and two plasmids containing SV40 sequences.
  • pL1 provides an SV40 early region promoter and an SV40 late region splice sequence.
  • pSV2-neo sequences provide a selectable marker for mammalian cell transformation and SV40 polyadenylation signal sequences.
  • pUC12 provides a multiple cloning site for cDNA insertion.
  • the plNG2003 vector has several useful restriction sites for modifications. These include a Hindlll site useful for the insertion of enhancer sequences, and a Hindlll to Xhol fragment useful for the insertion of alternate promoter sequences. This vector is useful in the expression of cDNA genes in mammalian cells.
  • Immunoglobulin enhancer elements have been shown to enhance transcription of genes in their vicinity in stably transformed mouse myeloma cells by several hundred fold (Gillies, S. D. et al., Cell, 33: 717, 1983; and Banerji, J. et al. Cell, 33: 729, 1983).
  • the mouse immunoglobulin heavy chain enhancer element was added to the cDNA expression vector plNG2003 ( Figure 10).
  • the mouse heavy chain enhancer region DNA was isolated from an M13 subclone of mouse heavy chain genomic DNA (M8-alpha-RX12, Deans, R. J., unpublished).
  • DNA isolated from a Sall plus EcoRl digestion of this subclone was modified with Hindlll linkers and inserted into the Hindlll site of pING2003, resulting in the new cDNA expression vector plNG2003E.
  • This vector is useful in the efficient expression of cDNA genes in mammalian cells, particularly mouse myeloma or hybridoma cell lines.
  • the cell line CRL8017 was obtained from the ATCC and subcloned. Subclones were grown and tested for mouse IgG anti-hepatitis B binding activity using a commercially available anti-HBsAg detection kit. Three positive subclones were found. Poly(A) + RNA was prepared from one of these subclones, and was fractionated on a methylmercury agarose gel. The RNA contained intact light chain and heavy chain mRNA's as inferred from specific hybridization to kappa UIG-MJK primer, and to the mouse heavy chain UIG-MJH3 probe (see Figure 7). In addition, the UIG-MJK primer was used for specific priming of anti-HBsAg poly(A) + RNA in a dideoxy sequencing reaction. Sufficient sequence was read to show that a major kappa RNA of the anti-HBsAg cell line contains the J K 2 sequence.
  • variable region cDNA synthesis were optimized by using heavy and light chain UIG primers on anti-HBsAg poly(A) + RNA.
  • Dideoxy chain extension experiments demonstrated that the mouse UIG-MJK primer and UIG-JH3 primer correctly primed kappa and heavy chain RNAs.
  • the reverse transcription was carried out in the absence of dideoxynucleotides, the main product using the kappa UIG-MJK primer was a 410 ⁇ 20 nucleotide fragment, while the main product using the heavy chain UIG-JH3 primer was a 430 ⁇ 30 nucleotide fragment. These correspond to the expected lengths of the variable and 5' untranslated regions of kappa and heavy chain immunoglobulin mRNAs.
  • the conditions for the optimal priming of poly(A) + RNA from CRL8017 cells should work well for poly(A) + RNA isolated from any cell line producing a monoclonal antibody.
  • oligonucleotides were designed and used for heavy chain V region cDNA synthesis. These two oligonucleotides are UIG-MJHBSTEII(13) and UIG-MJH3 ( Figures 7 and 8).
  • the primer sequence was designed to introduce a BstEll recognition site (GGTGACC) in the clone so that it could be joined at this site to the human IgGI constant module at the analogous position at the latter's J region. In this case, the primer had a single G to U mismatch with the mouse mRNA sequence that uses the J H 3 coding sequence.
  • the UIG-MJHBSTEII(13) primer was 13 bases long and the mismatched residue was flanked by 7 matches 5' and 5 matches 3' of it. This was the 13-mer BstEll primer.
  • a 21-mer primer specific for mouse J H 3 (UIG-MJH3) was used. This primer had a perfect match for the 17 nucleotides on its 3' end.
  • the first strand cDNA products made via the 13-mer BstEll and the 21-mer J H 3 primers included bands of approximately 430 nucleotides, which represented the entire V H region. Under the standard priming conditions used, the priming efficiency of the 13-mer BstEll was much less than that of the 21-mer J H 3. Accordingly, a cDNA library was generated from the first strand synthesis from each of these primers, using the method of Gubler and Hoffman, supra.
  • the 21-mer J H 3 library was screened with the 21-mer J H 3 oligonucleotide. Filter hybridization was done at 300, overnight, according to de Lange, T. et al., Cell, 34: 891-900 (1983). The filters were then washed at 51 ° in 6 x SSC, 0.1% SDS. Five colonies were selected. The largest had an insert of approximately 460 bp. More significantly, it contained three restriction sites predicted from the known J H 3 sequence, which are present upstream of the primer sequence. This clone, pJ3-11, was sequenced using the J H 3 primer by the chain-termination method (Wallace, R. B. et al., Gene, 16: 21-26 (1981)).
  • the sequence obtained has the remaining J H 3 coding segment.
  • a 13-nucleotide segment matched to a published D segment sequence (Dsp 2.2) (Kurosawa, Y. et al., J. Exp. Med., 155: 201 (1982), and Tonegawa, S., Nature, 302: 575 (1983)).
  • Dsp 2.2 published D segment sequence
  • a nonapeptide predicted from this area showed characteristic homology to the published mouse heavy chain V subgroups at amino acid residues 86 to 94, comprising the FR3 of heavy chain molecules.
  • Plasmid pJ3-11 represented a rearranged VDJ sequence, and apparently contained the anti-hepatitis V H sequence produced by the cell line.
  • V H cDNA clones were isolated from a cDNA library generated from the 21-mer oligonucleotide UIG-MJH3BSTEII primer containing a BstEll site. These clones may provide alternate V H cDNA sequences to join to human C H sequences.
  • the oligonucleotide UIG-JK2BGLII ( Figure 7B), was designed to introduce a Bglll site into the J K 2 region. Digestion with Bglll would then allow direct insertion of a V K cDNA coding region into the Bcll site of the previously noted human C K vector, p1NG2001. This insertion would result in the precise joining of a mouse variable region segment (including the J region) to a human kappa constant region segment, each in the proper coding frame and with no alteration in amino acid sequence for either mouse variable or human constant region.
  • the JK2BGLII oligonucleotide was used to prime anti-HBsAg mRNA to form a cDNA library as for heavy chain, supra, in pUC9.
  • the cDNA was size-selected by polyacrylamide gel electrophoresis prior to cloning, and 80% of the cDNA clones were shown to have insert sizes between 300 and 750 nucleotides in length.
  • Replica filters of this library were screened with two oligonucleotides, the original primer and a second probe complementary to J K 2 sequence 5' to the original primer.
  • the anti-hepatitis B monoclonal cell line CRL 8017 secretes immunoglobulins with at least two different light chains.
  • One of them is derived from the myeloma NS-1, which was used as a fusion partner in generating the anti-hepatitis B cell line.
  • NS-1 is derived from the myeloma MOPC21
  • MOPC21 V K mRNA may be present in the V K cDNA library from the antihepatitis monoclonal cell line.
  • one cDNA clone (p6D4B) analyzed has an identical restriction enzyme map to that of MOPC21 V K cDNA, except for the inserted Bglll site.
  • the first is that it is possible to effectively use an oligonucleotide to introduce a restriction enzyme site while cloning a V K region from a hybridoma cell line.
  • the second is that one must carefully monitor hybridoma cell lines for the presence of multiple V region sequences, only one of which is the desired sequence.
  • poly(A) + RNA was bound to nitrocellulose by the formaldehyde "Dot blot" procedure of White and Bancroft, J. Biol. Chem., 257: 8569 (1982).
  • the RNA was hybridized to 32 P-labeled oligonucleotide probes specific for each functional kappa J region. These probes are shown in Figure 7B as the UIG probes 5JK1, MJK, 5JK4, and 5JK5.
  • the results showed that the mRNA hybridized strongly to both MJK and 5JK4 oligonucleotide probes, indicating that both J K 2 and J K 4 sequences were present. Since J K 2 mRNA had been previously identified as the one derived from the parental hybridoma partner NS-1, it was concluded that the J K 4 mRNA encoded the anti-hepatitis binding specificity of the CRL 8017 cells.
  • JK4BGLII thee oligonucleotide primer
  • JK4BGLII thee oligonucleotide primer
  • the JK4BGLII primer was used to prime first strand cDNA synthesis to construct a cDNA library by the same method used to construct a JK2BGLII primed cDNA library, except that cDNA was not size selected prior to cloning.
  • Figure 7B tabulates the mismatches that each primer has with other functional mouse kappa J region sequences. Note that J K 4 has five mismatches in 21 nucleotides when compared with the JK2BGLII primer, and 3 in 23 with the JK4BGLII primer.
  • a new cDNA library was constructed by first priming with an oligonucleotide (5'-ATATTTGCT-GATGCT CT-3') complementary to pV17 sequences 155 bases from the 5' end. From this library, clones hybridizing to a pV17 DNA fragment probe were selected, and some of these new cDNA clones have the initiator ATG plus about 20 nucleotides of 5' untranslated region. One of these clones, p2-12, supplies a 5' untranslated region of 23 nucleotides and a complete ATG initiator codon. When p2-12 was combined with pV17 derived sequences, a variable region with a complete 5' end was formed (pING2013E).
  • an oligonucleotide 5'-ATATTTGCT-GATGCT CT-3'
  • V17-IVM site-directed mutagenesis on the existing light chain clone was used to simultaneously remove the poly-G tract and place a ribosome recognition sequence adjacent to the initiator ATG.
  • the Pstl fragment from pV17 was subcloned into M13mp18.
  • the resultant plasmid pV17-IVM provided an alternate mouse variable region for joining to human constant region modules.
  • the complete nucleotide sequence of the variable region from pV17 was then determined.
  • the sequence shows that pV17 contains a V K -J K junction region, containing several conserved amino acids, and the hybrid J K 2/J K 4 region formed by priming the J K 4 RNA with the UIG-JK2BGLII oligonucleotide.
  • the V K region in pV17 is non-functional, because the V K and J K regions are not in the same coding frame. Translation of the pV17 V region would thus result in an abnormal immunoglobulin light chain where the J region is translated in an incorrect frame.
  • This defect may be caused by aberrant V-J joining, resulting in a non-functional kappa mRNA, as has been observed by Kelley, D.E. et al., Mol. Cell. Biol., 5:1660-1675 (1985).
  • V K cDNA clones which are not from either of the two V K cDNA classes found so far (MOPC21-p6D4B, pV17).
  • a cDNA clone (p1E9L-81 ) that contains the kappa constant region, but has a different V K region than that of MOPC21 or pV17 was discovered.
  • This method of screening oligo-dT primed cDNA libraries is a useful alternative to oligonucleotide screening of cDNA libraries, because nick-translated probes of high specific activity are used. Also, this method allows the simultaneous isolation of several classes of V region clones, such as all V K clones, by appropriate probe choice.
  • the UIG-JK2BGLII-primed cDNA library made from CRL 8017 RNA was screened with the UIG-5JK2 oligonucleotide probe (see Figure 7). A new class of V K cDNA clones was found whose members are homologous to p1 E9L-81 and hybridize to the UIG-5JK2 probe, but not to a MOPC21 V K probe.
  • restriction endonuclease site maps and nucleotide sequences of these clones also differ from MOPC21-homologous V K cDNA clones from CRL8017 cells. These clones, however, have an aberrant V-J joint which results in a nonfunctional mRNA, and appear to be identical to one described by Cabilly and Riggs (Gene, 40:157 (1985)).
  • the anti-hepatitis B cell line CRL8017 has at least three classes of V K mRNA corresponding to the above described cDNA clones p6D4B (MOPC21), p1E9L, and pV17.
  • the plE9L and pV17 clones are derived from mRNA from aberrantly rearranged Kappa genes, while the p6D4B clone is derived from the parent hybridoma fusion partner NS-1. None of these clones appear to encode the desired anti-hepatitis light chain.
  • V region sequences in pMVHCa-13 were joined to the human IgGI constant (C) region clone pGMH-6. Due to the presence of a second BstEll site within the IgGI CH1 region of pGMH-6, a multi-step ligation was required. First, the 220 nucleotide BstEll fragment from the J-CH1 region of pGMH-6 was ligated to the 1100 nucleotide IgG region BstEll to BamHl fragment of pGMH-6.
  • V region of the hybrid heavy chain gene in pMVHCc-24 was further analyzed by partial sequence analysis. This analysis showed that the cloned V region contained a D sequence which matches a known D sequence, DSP2.2 (Kurosawa and Tonegawa, supra). The sequence also predicted a 19 amino acid leader peptide similar to known mouse V heavy chain leader peptide sequences, and a 5' untranslated region of at least 3 nucleotides.
  • the BamHl fragment containing the mouse-human hybrid heavy chain gene of pMVHCc-24 was cloned into BamHl digested plNG2003E vector, resulting in the expression plasmid plNG2006E ( Figure 11).
  • the plNG2006E plasmid should have an increased probability of efficient expression of the mouse-human chimeric immunoglobulin gene in B lymphoid cells because of the presence of the mouse heavy chain enhancer region.
  • a modification of the chimeric heavy chain gene present in pMVHCc-24 was done to provide an alternate heavy chain gene which lacks the oligo-dC region preceding the initiator ATG.
  • the plNG2012E and plNG2006E vectors are identical except for the nucleotides immediately preceding the ATG, as shown in Figure 12.
  • Bacteria harboring the plNG2006E and pSV2-neo plasmids were converted into protoplasts by the method of Sandri-Goldin, R. M. et al., Mol. Cell. Biol., 1: 743 (1981).
  • the protoplasts were then separately fused to SP2/0-Ag14 hybridoma cells (ATCC CRL 1581) by treatment with polyethyleneglycol (Ochi, A. et al., Nature, 302: 340, 1983).
  • the fused cells were allowed to recover for 72 hours in complete medium before plating at 10,000 or 50,000 cells per well in a 96-well tissue culture plate.
  • the cells were selected with G418 at 0.8 mg/ml for two weeks, when growth in some wells was clearly evident.
  • RNA and protein level were tested for immunoglobulin gene expression at the RNA and protein level.
  • Total cell RNA was prepared from transfected cells, bound to nitrocellulose and hybridized to nick-translated probes specific for the mouse-human hybrid heavy chain gene. Two clones were found which have a strong signal, representing expression of the gene at the RNA level. The amount of total cellular RNA hybridizing to the mouse-human probe appeared to be approximately 1/10 the level of heavy chain RNA in the original hybridoma cells. This probably represented about 1 % of the total mRNA of the transfected cell.
  • the transfected mouse cells were also tested for production of cytoplasmic human heavy chain protein by an ELISA assay. It was found that 3 out of 7 plNG2006E transfected cell lines produced detectable levels of human heavy chain protein.
  • the mouse cell transformant producing the most mouse-human heavy chain protein gave a signal in the ELISA assay comparable to that of a 1/100 dilution of a human B cell line producing intact human immunoglobulin IgGI. This modest level of detected mouse-human heavy chain protein may be due to several factors, including instability of heavy chains in the absence of light chains in hybridoma cells, or incorrect processing of the chimeric gene transcript.
  • EXAMPLE III A Human-Mouse Chimeric Antibody with Cancer Antigen Specificity
  • L6 monoclonal antibody was obtained from a mouse which had been immunized with cells from a human lung carcinoma, after which spleen cells were hybridized with NS-1 mouse myeloma cells.
  • the antibody binds to a previously not identified carbohydrate antigen which is expressed in large amounts at the surface of cells from most human carcinomas, including lung carcinomas (adeno, squamous), breast carcinomas, colon carcinomas and ovarian carcinomas, while the antigen is only present at trace levels in normal cells from the adult host.
  • MAb L6 is an IgG2a and can mediate antibody dependent cellular cytotoxicity, ADCC, in the presence of human peripheral blood leukocytes as a source of effector cells, so as to lyse L6 positive tumor cells, and it can lyse L6 positive tumor cells in the presence of human serum as a source of complement; the lysis is detected as the release of 51Cr from labelled cells over a 4 hour incubation period.
  • MAb L6 can localize to L6 positive tumors xenotransplanted onto nude mice, and it can inhibit the outgrowth of such tumors.
  • MAb L6 is described in Cancer Res. 46:3917-3923, 1986 (on MAb specificity) and in Proc. Natl. Acad. Sci. 83:7059-7063, 1986 (on MAb function)
  • Frozen cells were thawed on ice for 10 minutes and then at room temperature. The suspension was diluted with 15 ml PBS and the cells were centrifuged down. They were resuspended, after washes in PBS, in 16 ml 3M LiCI, 6M urea and disrupted in a polytron shear. The preparation of mRNA and the selection of the poly(A+) fraction were carried out according to Auffray, C. and Rougeon, F., Eur. J. Biochem. 107:303, 1980.
  • the poly (A+) RNA from L6 was hybridized individually with labeled J H 1 ⁇ J H 2, J H 3 and J H 4 oligonucleotides under conditions described by Nobrega et al. Anal. Biochem 131:141, 1983). The products were then subjected to electrophoresis in a 1.7% agarose-TBE gel. The gel was fixed in 10% TCA, blotted dry and exposed for autoradiography. The result showed that the L6 v H contains J H 2 sequences.
  • V K mRNA For the analysis of the V K mRNA, the dot-blot method of White and Bancroft J. Biol. Chem. 257:8569, (1982) was used. Poly (A+) RNA was immobilized on nitrocellulose filters and was hybridized to labeled probe-oligonucleotides at 40 ° in 4xSSC. These experiments show that L6 contains J K 5 sequences. A faint hybridization to J K 2 was observed.
  • a library primed by oligo (dT) on L6 poly (A+) RNA was screened for kappa clones with a mouse C K region probe. From the L6 library, several clones were isolated. A second screen with a 5' J K 5 specific probe identified the L6 (J K 5) light-chain clones. Heavy chain clones of L6 were isolated by screening with the J H 2 oligonucleotide.
  • the heavy and light chain genes or gene fragments from the cDNA clones, pH3-6a and pL3-12a were inserted into M13 bacteriophage vectors for nucleotide sequence analysis.
  • the complete nucleotide sequences of the variable region of these clones were determined (FIGURES 15 and 16) by the dideoxy chain termination method. These sequences predict V region amino acid compositions that agree well with the observed compositions, and predict peptide sequences which have been verified by direct amino acid sequencing of portions of the V regions.
  • the nucleotide sequences of the cDNA clones show that they are immunoglobulin V region clones as they contain amino acid residues diagnostic of V domains (Kabat et al., Sequences of Proteins of Immunological Interest; U.S. Dept of HHS, 1983).
  • the L6 V H belongs to subgroup II.
  • the cDNA predicts an N-terminal sequence of 24 amino acid residues identical to that of a known V H (45-165 CRI; Margolies et al. Mol. Immunol. 18:1065, 1981).
  • the L6 V H has the J H 2 sequence.
  • the L6 V L is from the V K -Kpnl family (Nishi et al. Proc. Nat. Acd. Sci. USA 82:6399, 1985), and uses J K 5.
  • the cloned L6 V L predicts an amino acid sequence which was confirmed by amino acid sequencing of peptides from the L6 light chain corresponding to residues 18-40 and 80-96.
  • the J-region mutagenesis primer J K Hindlll as shown in FIGURE 17B, was utilized.
  • oligo (dC) sequence upstream of the ATG codon in a V H chimericgene interferes with proper splicing in one particular gene construct. It was estimated that perhaps as much as 70% of the RNA transcripts had undergone the mis-splicing, wherein a cryptic 3' splice acceptor in the leader sequence was used. Therefore the oligo (dC) sequence upstream of the initiator ATG was removed in all of the clones.
  • an oligonucleotide was used which contains a Sall restriction site to mutagenize the L6 V K clone.
  • the primer used for this oligonucleotide-directed mutagenesis is a 22-mer which introduces a Sall site between the oligo (dC) and the initiator met codon (FIGURE 19).
  • nuclease BAL-31 was used to chew away the oligo (dC) in the L6 V H clone pH3-6a.
  • the size of the deletion in two of the mutants obtained was determined by nucleotide sequencing and is shown in FIGURE 17. In both of these mutuants (delta 4 and delta 21), all of the oligo (dC) 5' to the coding region were deleted.
  • the human C gamma 1 gene module is a cDNA derived from GM2146 cells (Human Genetic Mutant Cell Repository, Newark, New Jersey). This C gamma 1 gene module was previously combined with a mouse V H gene module to form the chimeric expression plasmid p1NG2012E.
  • L6 chimeric heavy chain expression plasmids were derived from the replacement of the V H module pING2012E with the V H modules of mutants delta 21 and delta 4 to give the expression plasmids pING2111 and plNG2112 (FIGURE 17). These plasmids direct the synthesis of chimeric L6 heavy chain when transfected into mammalian cells.
  • the Sall to Hindlll fragment of the mouse V K module was joined to the human C K module by the procedure outlined in FIGURE 18, forming p1NG2119.
  • Replacement of the neo sequence with the E. coli gpt gene derived from pSV2-gpt resulted in pING2120, which expressed L6 chimeric light chain and confers mycophenolic acid resistance when transfected into mammalian cells.
  • both heavy and light chain chimeric genes in the same plasmid allows for the introduction into transfected cells of a 1:1 gene ratio of heavy and light chain genes leading to a balanced gene dosage. This may improve expression and decrease manipulations of transfected cells for optimal chimeric antibody expression.
  • the DNA fragments derived from the chimeric heavy and light chain genes of plNG2111 and plNG2119 were combined into the expression plasmid plNG2114 (FIGURE 19).
  • This expression plasmid contains a selectable neo R marker and separate transcription units for each chimeric gene, each including a mouse heavy chain enhancer.
  • Electroporation was used (Potter et al. supra; Toneguzzo et al. Mol. Cell Biol. 6:703 1986) for the introduction of L6 chimeric expression plasmid DNA into mouse Sp2/0 cells.
  • the electroporation technique gave a transfection frequency of 1-10 x 10- 5 for the Sp2/0 cells.
  • the two gene expression plasmid plNG2114 was linearized by digestion with Aatll restriction endonuclease and transfected into Sp2/0 cells, giving approximately fifty G418 resistant clones which were screened for human heavy and light chain synthesis.
  • the levels of chimeric antibody chain synthesis from the two producers, D7 and 3E3, are shown in Table 3.
  • Chimeric L6 antibody was prepared by culturing the D7 transfectant cells for 24 hours at 2x10 6 cells/ml in 5 1 DMEM supplemented with HEPES buffer and penicillin and streptomycin. The supernatant was concentrated over an Amicon YM30@ membrane in 10mM sodium phosphate buffer, pH8.0.
  • the preparation was loaded over a DEAE-Cellulose column, which separated the immunoglobulin into unbound and bound fractions.
  • Samples from the DEAE-unbound, DEAE- bound and the pre-DEAE preparations (from 1.6 ul of medium) was separately purified by affinity chromatography on a Protein-A Sepharose@ column, eluting with 0.1 M sodium citrate,
  • the antibody can be further concentrated by steps j. and k. of the previous procedure.
  • the samples were tested with a binding assay, in which cells of both an L6 antigen-positive and an L6 antigen-negative cell line were incubated with standard mouse monoclonal antibody L6, chimeric L6 antibody derived from the cell culture supernatants, and chimeric L6 antibody derived from ascites (as previously described) followed by a second reagent, fluorescein-isothiocyanate (FITC)-conjugated goat antibodies to human (or mouse, for the standard) immunoglobulin.
  • FITC fluorescein-isothiocyanate
  • the binding assay showed strong reactivity of the chimeric L6 on the L6 antigen positive cell line and total lack of reactivity on the negative cell line
  • the next step was to test for the ability of the chimeric L6 to inhibit the binding of mouse L6 to antigen positive cells; such inhibition assays are used routinely to establish the identity of two antibodies' recognition of antigen. These data are discussed below ("Inhibition of binding"). As part of these studies, a rough estimate of antibody avidity was made.
  • Binding Assays Cells from a human colon carcinoma line, 3347, which had been previously shown to express approximately 5 x 10 5 molecules of the L6 antigen at the cell surface, were used as targets. Cells from the T cell line HSB2 was used as a negative control, since they, according to previous testing, do not express detectable amounts of the L6 antigen. The target cells were first incubated for 30 min at 4 ° C with either the chimeric L6 or with mouse L6 standard, which had been purified from mouse ascites.
  • both the chimeric and the mouse standard L6 bound significantly, and to approximately the same extent, to the L6 positive 3347 line. They did not bind above background to the L6 negative HSB2 line.
  • Both the chimeric and mouse standard L6 inhibited the binding of the directly labelled L6 antibody, with the binding curves being parallel.
  • the chimeric antibody was slightly less effective than the standard, as indicated by the results which showed that 3.4 ug/ml of the pooled chimeric L6 MAb, as compared to 2.0 ug/ml of the standard mouse L6 MAb was needed for 50% inhibition of the binding, and that 5.5 ug/ml of the chimeric L6 (derived from ascites) as compared to 2.7 ug/ml of the standard mouse L6 MAb was needed for 50% inhibition of binding.
  • the chimeric L6 was superior to the simultaneously tested sample of mouse L6 in causing ADCC, as measured by a 4 hr 51 Cr release test.
  • Tables 6 and 6A-6B present the data from studies on complement-mediated target cell lysis. In this case, a high cytolytic activity was observed with both the mouse and the chimeric L6 antibodies.
  • the chimeric L6 antibody binds to L6 antigen positive tumor cells to approximately the same extent as the mouse L6 standard and with approximately the same avidity. This is significant for the following reasons: the L6 antibody defines (a) a surface carbohydrate antigen, and (b) a protein antigen of about 20,000 daltons, each of which is characteristic of non-small cell lung carcinoma (NSCLC) and certain other human carcinomas. Significantly, the L6 antibody does not bind detectably to normal cells such as fibroblasts, endothelial cells, or epithelial cells in the major organs. Thus the chimeric L6 monoclonal antibody defines an antigen that is specific for carcinoma cells and not normal cells.
  • NSCLC non-small cell lung carcinoma
  • the chimeric L6 exerts profound biological effects upon binding to its target, which make the chimeric antibody a prime candidate for tumor immunotherapy.
  • the results presented herein demonstrate that chimeric L6 is capable of binding to tumor cells and upon binding kills the tumor cells, either by ADCC or CDC. Such tumor killing activity was demonstrated using concentrations of chimeric L6 antibody as low as 0.01 ug/ml (10ng/ml).
  • the chimeric L6 monoclonal antibodies will be advantageously used not only for therapy with unmodified chimeric antibodies, but also for development of various immunoconjugates with drugs, toxins, immunomodulators, isotopes, etc., as well as for diagnostic purposes such as in vivo imaging of tumors using appropriately labelled chimeric L6 antibodies.
  • immunoconjugation techniques are known to those skilled in the art and can be used to modify the chimeric L6 antibody molecules of the present invention.
  • transfected hybridoma C255 (corresponds to 3E3 cells, supra), ATCC HB 9240 and transfected hybridoma C256 (C7 cells, supra), ATCC HB 9241.
  • EXAMPLE IV A Human-Mouse Chimeric Antibody with Specificity for Human B-Cell Antigen
  • the 2H7 mouse monoclonal antibody (gamma 2b K ) recognizes a human B-cell surface antigen, Bp35 (Clark, E A. et al., Proc. Nat. Acad. Sci. USA, 82:1766 (1985)).
  • Bp35 a human B-cell surface antigen
  • the Bp35 molecule plays a role in B-cell activation.
  • mRNA was prepared from the 2H7 cell line. Two cDNA libraries were generated - one using the heavy chain UIG-H primer and the other, oligo(dT). One V H clone, pH2-11, was isolated upon screening with the same UIG-H oligonucleotide.
  • a mouse kappa-specific DNA fragment was used to screen the oligo(dT) library.
  • Candidate clones were further screened with a mouse J K 5 sequences.
  • the light chain UIG-K was then used to engineer a restriction enzyme site in the J region.
  • the two cDNA clones were also modified at the 5' end to remove the artificial oligo d[C] sequence.
  • pH2-11 this was carried out by using the restriction enzyme Ncol which cuts one nucleotide residue 5' of the ATG initiator codon.
  • pL2-12 this was achieved by an oligonucleotide in vitro mutagenesis using a 22-mer containing a Sall site.
  • the DNA sequences of these two clones are shown in Figures 21, 22.
  • the V H module was joined to the human C gamma 1 module (pGMH6) at the J H BstEll site, and to construct the chimeric light chain the V K module was joined to the human C K module (pGML60) at the J K Hindlll site.
  • the expression vector sequences were derived from pING2012-neo as well as pING2016-gpt.
  • the constructed plasmids are plNG2101 (V H C gamma 1-neo).
  • plNG2106 V K C K -neo
  • plNG2107 V K C K -gpt).
  • plNG2101 and plNG2106 were also used to generate plasmids containing both genes. They are pHL2-11 and pHL2-26.
  • pING2106 and pING2014 were combined to a two light-chain plasmid, pLL2-25, to compensate for the poorer (compared to heavy chain) steady-state accumulation of light chain protein in transfected cells.
  • Fig. 24 shows the changes made to the variable region sequences during the construction.
  • the plasmid, pHL2-11, was linearized by Aatll; and the DNA was used to transfect Sp2/0 cells by electroporation. Transformants were selected in G418-DMEM.
  • One transformant, 1 C9 produces 9.3 ng/ml chimeric kappa and 33-72 ng/ml chimeric gamma 1 protein as assayed by ELISA. Southern analysis of 1 C9 DNA showed that there is one copy of the plasmid integrated in Sp2/0 genome.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Plant Pathology (AREA)
  • Mycology (AREA)
  • Cell Biology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Claims (38)

1. Vektor, umfassend eine cDNA-Sequenz, die für die vollständige variable Region einer Immunglobulinkette kodiert, wobei diese Kette eine vollständige V-J-Verbindung im Fall einer leichten Kette und eine vollständige V-D-J-Verbindung im Fall einer schweren Kette enthält, und wobei dem Vektor eine konstante Region oder Intronsequenzen fehlen.
2. DNA-Fragment, umfassend eine cDNA-Sequenz, die für die vollständige konstante IgG1-Region für eine schwere Human-Immunglobulinkette kodiert, wobei dem Fragment eine variable Region fehlt und der konstanten Region Intronsequenzen fehlen.
3. Vektor, umfassend eine kontinuierliche kodierende Sequenz, die nicht von Introns unterbrochen wird, umfassend:
i) eine für die variable Region einer nicht-Human-Immunglobulinkette kodierende DNA-Sequenz einschließlich einer V-J-Verbindung im Fall einer leichten Kette und einer V-D-J-Verbindung im Fall einer schweren Kette;
ii) eine für die konstante Region einer Human-Immunglobulinkette kodierende DNA-Sequenz.
4. Vektor nach Anspruch 1 oder 3, der ein Plasmid ist.
5. Bakterium, transformiert mit dem Vektor oder dem Fragment nach einem der Ansprüche 1, 2, 3 oder 4.
6. Säugetierzelle oder Hefe, transfiziert mit dem Vektor oder Fragment nach einem der Ansprüche 1, 2, 3 oder 4.
7. Polynukleotidmolekül, umfassend eine Konsensussequenz für die J-Region eines Immunglobulinmoleküls für eine schwere Kette, wobei die Konsensussequenz mindestens 80% Sequenzhomologie mit bekannten J-Region-Sequenzen aufweist, ohne damit identisch zu sein.
8. Molekül nach Anspruch 7, wobei die Sequenz für die J-Region einer schweren Human-Kette ist.
9. Molekül nach Anspruch 7, wobei die Sequenz für die J-Region einer schweren Maus-Kette ist.
10. Polynukleotidmolekül, umfassend eine Konsensussequenz für die J-Region eines Immunglobulinmoleküls für eine leichte Kette, wobei die Konsensussequenz mindestens 80% Sequenzhomologie mit bekannten J-Region-Sequenzen aufweist, ohne damit identisch zu sein.
11. Molekül nach Anspruch 10, wobei die Sequenz für eine Human-Kappa-J-Region ist.
12. Molekül nach Anspruch 10, wobei die Sequenz für eine Maus-Kappa-J-Region ist.
13. Molekül nach Anspruch 10, wobei die Sequenz für eine Maus-Lambda-J-Region ist.
14. Verfahren zum direkten Herstellen einer genetischen Sequenz, die für eine chimäre Immunglobulinkette mit einer konstanten Human-Region und einer variablen nicht-Human-Region jeder gewünschten Spezifität kodiert, umfassend:
a) das Bereitstellen einer für eine vollständige variable nicht-Human-Region kodierenden cDNA-Sequenz einschließlich einer vollständigen V-J-Verbindung im Fall einer leichten Kette und einer vollständigen V-D-J-Verbindung im Fall einer schweren Kette, wobei Sequenzen eines konstanten Bereiches fehlen;
b) das Bereitstellen eines Vektors, der eine genetische Sequenz enthält, die für die konstante Region kodiert;
c) das funktionelle Verbinden der Sequenz a) mit dem Vektor b).
15. Verfahren nach Anspruch 14, wobei Schritt (c) das funktionelle Verbinden der cDNA-Sequenz mit der Sequenz aus Schritt (c) in einem Plasmid umfaßt.
16. Verfahren nach Anspruch 15, das weiter das Transformieren des Plasmides in einen Wirt umfaßt, der zur Expression des Plasmides fähig ist.
17. Verfahren nach einem der Ansprüche 14 bis 16, wobei die Kette eine schwere Kette ist.
18. Verfahren nach einem der Ansprüche 14 bis 16, wobei die Kette eine leichte Kette ist.
19. Verfahren nach Anspruch 14, wobei der Schritt (a) umfaßt:
a') das Bereitstellen einer mRNA, die für die variable Region kodiert, aus einer Zelle, die monoklonale Antikörper der gewünschten Spezifität ausscheidet; und
a") das Starten (Priming) der Bildung einer davon abgeleiteten cDNA durch reverse Transkription unter Verwendung der mRNA als Matrize mit einem Polynukleotidmolekül, das eine genetische Konsensussequenz für die J-Region der Immunglobulinkette umfaßt, wobei die Konsensussequenz mindestens 80% Sequenzhomologie mit bekannten J-Region-Sequenzen aufweist.
20. Verfahren nach Anspruch 19, wobei die genetische Konsensussequenz aus der Gruppe ausgewählt ist, die besteht aus:
(i) Human-J-Region für schwere Ketten;
(ii) Maus-J-Region für schwere Ketten;
(iii) Human-Kappa-J-Region;
(iv) Maus-Kappa-J-Region; und
(v) Maus-Lambda-J-Region.
21. Verfahren nach Anspruch 19, wobei die genetische Konsensussequenz aus der Gruppe ausgewählt ist, die aus den mit MJH1, MJH2, MJH3, MJH3-BSTEII, MJH-BSTEII(13), MJH4, 5JK1, 5JK2, JK2BGLII, 5JK4, JK4BGLII, 5JK5 und MJK bezeichneten besteht:
Figure imgb0021
22. Verfahren nach Anspruch 19, wobei die Konsensussequenz weiter die für die Erkennungsstelle eines Restriktionsendonukleaseenzymes kodierende Sequenz umfaßt.
23. cDNA-Expressionsvektoren mit Restriktionsendonuklease-Schnittstellenkarten, wie in Figur 10 gezeigt, die einen Promotor aus der frühen Region von SV40, eine Spleißsequenz aus der späten Region von SV40, den selektierbaren Marker neo, SV40 polyA-Signalsequenzen, eine Vielfachklonierungstelle (pING2003) und gegebenenfalls zusätzlich ein Enhancer-Element aus einer schweren Mauskette (pING2003 E) enthalten.
24. Verfahren nach Anspruch 19, wobei die genetische Konsensussequenz aus der Gruppe ausgewählt ist, die aus den als UIGH, UIGK und MJH2-Apal bezeichneten besteht:
Figure imgb0022
EP86906676A 1985-11-01 1986-10-27 Modulare einheit von antikörpergenen, daraus hergestellte antikörper und verwendung Revoked EP0247091B1 (de)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AT86906676T ATE95235T1 (de) 1985-11-01 1986-10-27 Modulare einheit von antikoerpergenen, daraus hergestellte antikoerper und verwendung.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US79398085A 1985-11-01 1985-11-01
US793980 1985-11-01

Related Child Applications (1)

Application Number Title Priority Date Filing Date
EP92115754.1 Division-Into 1992-09-15

Publications (3)

Publication Number Publication Date
EP0247091A4 EP0247091A4 (de) 1987-10-27
EP0247091A1 EP0247091A1 (de) 1987-12-02
EP0247091B1 true EP0247091B1 (de) 1993-09-29

Family

ID=25161322

Family Applications (1)

Application Number Title Priority Date Filing Date
EP86906676A Revoked EP0247091B1 (de) 1985-11-01 1986-10-27 Modulare einheit von antikörpergenen, daraus hergestellte antikörper und verwendung

Country Status (6)

Country Link
EP (1) EP0247091B1 (de)
JP (1) JPS63501765A (de)
AU (1) AU606320B2 (de)
DE (1) DE3689123T2 (de)
DK (1) DK175680B1 (de)
WO (1) WO1987002671A1 (de)

Families Citing this family (488)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4816567A (en) * 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US5576195A (en) * 1985-11-01 1996-11-19 Xoma Corporation Vectors with pectate lyase signal sequence
US5618920A (en) * 1985-11-01 1997-04-08 Xoma Corporation Modular assembly of antibody genes, antibodies prepared thereby and use
EP0256654B1 (de) * 1986-07-07 1996-09-18 Centocor, Inc. Chimärisches Murine-Mensch-Immunoglobulin, spezifisch für tumorassoziertes 17-1A Antigen
IL84285A (en) * 1986-10-27 1993-03-15 Int Genetic Engineering Chimeric antibody with specificity to human tumor antigen
US6893625B1 (en) 1986-10-27 2005-05-17 Royalty Pharma Finance Trust Chimeric antibody with specificity to human B cell surface antigen
IL85035A0 (en) * 1987-01-08 1988-06-30 Int Genetic Eng Polynucleotide molecule,a chimeric antibody with specificity for human b cell surface antigen,a process for the preparation and methods utilizing the same
US5258498A (en) * 1987-05-21 1993-11-02 Creative Biomolecules, Inc. Polypeptide linkers for production of biosynthetic proteins
DE3856559T2 (de) * 1987-05-21 2004-04-29 Micromet Ag Multifunktionelle Proteine mit vorbestimmter Zielsetzung
US5132405A (en) * 1987-05-21 1992-07-21 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US5091513A (en) * 1987-05-21 1992-02-25 Creative Biomolecules, Inc. Biosynthetic antibody binding sites
US6657050B1 (en) 1987-05-29 2003-12-02 Tanox, Inc. Chimeric viral-neutralizing immunoglobulins
US5834599A (en) * 1987-05-29 1998-11-10 Tanox Biosystems, Inc. Immunoconjugates which neutralize HIV-1 infection
US5981278A (en) * 1987-05-29 1999-11-09 Tanox, Inc. Chimeric monoclonal antibodies which neutralize HIV-1 infection and their applications in therapy and prevention for AIDS
CA1341235C (en) * 1987-07-24 2001-05-22 Randy R. Robinson Modular assembly of antibody genes, antibodies prepared thereby and use
GB8720833D0 (en) * 1987-09-04 1987-10-14 Celltech Ltd Recombinant dna product
NZ226694A (en) * 1987-10-28 1994-04-27 Oncogen Human immunoglobulin produced by recombinant dna techniques
US4978745A (en) * 1987-11-23 1990-12-18 Centocor, Inc. Immunoreactive heterochain antibodies
US5843708A (en) * 1988-01-05 1998-12-01 Ciba-Geigy Corporation Chimeric antibodies
GB8800077D0 (en) * 1988-01-05 1988-02-10 Ciba Geigy Ag Novel chimeric antibodies
WO1989007142A1 (en) * 1988-02-05 1989-08-10 Morrison Sherie L Domain-modified constant region antibodies
WO1989011533A1 (en) * 1988-05-23 1989-11-30 The United States Of America, As Represented By Th Cloned gene for expression of antibodies reacting with human ovarian cancer
US5576184A (en) * 1988-09-06 1996-11-19 Xoma Corporation Production of chimeric mouse-human antibodies with specificity to human tumor antigens
DE68929167T2 (de) * 1988-09-06 2000-11-16 Xoma Corp., Berkeley Genexpressions-Elemente und Herstellung von chimären Maus-Mensch-Antikörpern
AU634186B2 (en) * 1988-11-11 1993-02-18 Medical Research Council Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors
US5530101A (en) * 1988-12-28 1996-06-25 Protein Design Labs, Inc. Humanized immunoglobulins
AU632843B2 (en) * 1989-08-10 1993-01-14 Juridical Foundation The Chemo-Sero-Therapeutic Research Institute Cat-mouse heterohybridoma and gene fragment coding for constant region of feline immunoglobulin
US5698426A (en) * 1990-09-28 1997-12-16 Ixsys, Incorporated Surface expression libraries of heteromeric receptors
US6893845B1 (en) 1990-09-28 2005-05-17 Applied Molecular Evolution, Inc. Surface expression libraries of heteromeric receptors
US5871974A (en) * 1990-09-28 1999-02-16 Ixsys Inc. Surface expression libraries of heteromeric receptors
ES2236682T5 (es) * 1991-01-21 2011-03-31 Elan Pharmaceuticals, Inc. Ensayo y modelo para la enfermedad de alzheimer.
US6797492B2 (en) 1991-05-17 2004-09-28 Merck & Co., Inc. Method for reducing the immunogenicity of antibody variable domains
US6800738B1 (en) 1991-06-14 2004-10-05 Genentech, Inc. Method for making humanized antibodies
EP0940468A1 (de) * 1991-06-14 1999-09-08 Genentech, Inc. Änderliches Gebiet einer humanisierter Antikörper
WO1994004679A1 (en) * 1991-06-14 1994-03-03 Genentech, Inc. Method for making humanized antibodies
US6129914A (en) 1992-03-27 2000-10-10 Protein Design Labs, Inc. Bispecific antibody effective to treat B-cell lymphoma and cell line
US7754211B2 (en) 1992-04-10 2010-07-13 Research Development Foundation Immunotoxins directed against c-erbB-2(HER-2/neu) related surface antigens
US5604102A (en) * 1992-04-15 1997-02-18 Athena Neurosciences, Inc. Methods of screening for β-amyloid peptide production inhibitors
US5441870A (en) * 1992-04-15 1995-08-15 Athena Neurosciences, Inc. Methods for monitoring cellular processing of β-amyloid precursor protein
US5851787A (en) * 1992-04-20 1998-12-22 The General Hospital Corporation Nucleic acid encoding amyloid precursor-like protein and uses thereof
US6610493B1 (en) 1993-06-17 2003-08-26 Brigham And Women's Hospital Screening compounds for the ability to alter the production of amyloid-β peptide
US6270766B1 (en) 1992-10-08 2001-08-07 The Kennedy Institute Of Rheumatology Anti-TNF antibodies and methotrexate in the treatment of arthritis and crohn's disease
US5605811A (en) * 1992-10-26 1997-02-25 Athena Neurosciences, Inc. Methods and compositions for monitoring cellular processing of beta-amyloid precursor protein
PL174721B1 (pl) 1992-11-13 1998-09-30 Idec Pharma Corp Przeciwciało monoklonalne anty-CD20
US7744877B2 (en) 1992-11-13 2010-06-29 Biogen Idec Inc. Expression and use of anti-CD20 Antibodies
ES2155099T3 (es) 1993-10-27 2001-05-01 Elan Pharm Inc Animales transgenicos que albergan alelos app que presentan una mutacion sueca.
CU22615A1 (es) 1994-06-30 2000-02-10 Centro Inmunologia Molecular Procedimiento de obtención de anticuerpos monoclonales murinos menos inmunogénicos. anticuerpos monoclonales obtenidos
US6576236B1 (en) 1994-07-01 2003-06-10 Dana Farber Cancer Institute Methods for stimulating T cell responses by manipulating a common cytokine receptor γ chain
ATE407207T1 (de) 1994-11-02 2008-09-15 Trophix Pharm Inc Natrium-ionenkanäle spezifisch für das periphäre nervensystem, dna welche diese kodiert, wirkstoff-suche und methoden zu deren herstellung und verwendung
IL114615A0 (en) 1995-07-16 1995-11-27 Yeda Res & Dev Modulators of the function of fas receptors and other proteins
US6717031B2 (en) 1995-06-07 2004-04-06 Kate Dora Games Method for selecting a transgenic mouse model of alzheimer's disease
US6248555B1 (en) 1995-08-31 2001-06-19 The General Hospital Corporation Genetic alterations related to familial alzheimer's disease
US7888466B2 (en) 1996-01-11 2011-02-15 Human Genome Sciences, Inc. Human G-protein chemokine receptor HSATU68
DK0961831T3 (da) 1996-08-16 2009-01-12 Human Genome Sciences Inc Humant alfa-endokin
US6653104B2 (en) 1996-10-17 2003-11-25 Immunomedics, Inc. Immunotoxins, comprising an internalizing antibody, directed against malignant and normal cells
CA2665133A1 (en) 1996-10-25 1998-05-07 Human Genome Sciences, Inc. Neutrokine .alpha.
ES2402947T3 (es) 1997-04-10 2013-05-10 Stichting Katholieke Universiteit University Medical Centre Nijmegen PCA3, genes PCA3 y métodos de uso
WO1998049313A2 (en) 1997-04-25 1998-11-05 Aquila Biopharmaceuticals, Inc. Characterization of granulocytic ehrlichia and methods of use
CA2288232A1 (en) 1997-05-02 1998-11-12 The Government Of The United States, Represented By The Secretary, Depar Tment Of Health And Human Services Immunotoxins directed against malignant cells
JP2002512521A (ja) 1997-05-30 2002-04-23 ヒューマン ジノーム サイエンシーズ，インコーポレイテッド ３２個のヒト分泌タンパク質
CA2206774A1 (en) 1997-06-16 1998-12-16 Rick E. Preddie "prionins", highly specific markers for noninvasive presymptomatic defection of tse diseases, and targets for therapeutic reagents to prevent and control tse diseases in animals and humans
DE69841140D1 (de) 1997-09-17 2009-10-22 Human Genome Sciences Inc Interleukin-17 rezeptor-ähnliches protein
DE69839060T2 (de) 1997-11-03 2009-01-15 Human Genome Sciences, Inc. Vegi, ein inhibitor der angiogenese und des tumorwachstums
JP2001524308A (ja) 1997-11-21 2001-12-04 ヒューマン ジノーム サイエンシーズ， インコーポレイテッド ケモカインα−５
US7179892B2 (en) 2000-12-06 2007-02-20 Neuralab Limited Humanized antibodies that recognize beta amyloid peptide
TWI239847B (en) 1997-12-02 2005-09-21 Elan Pharm Inc N-terminal fragment of Abeta peptide and an adjuvant for preventing and treating amyloidogenic disease
EP1093457B8 (de) 1998-03-19 2011-02-02 Human Genome Sciences, Inc. Der gemeinsamen gamma-kette ähnlicher zytokinrezeptor
CZ303898B6 (cs) 1998-08-11 2013-06-19 Biogen Idec Inc. Rituximab jako lécivo pro udrzovací terapii
GB9822763D0 (en) * 1998-10-20 1998-12-16 Univ Sheffield Immunoglobin variant
EP1131093A4 (de) 1998-11-09 2002-05-02 Idec Pharma Corp Behandlung hämatologischer bösartigkeiten assoziiert mit zirkulierenden tumorzellen unter verwendung chimärer anti-cd20 antikörper
AU764211C (en) 1998-12-01 2006-03-30 Abbvie Biotherapeutics Inc. Humanized antibodies to gamma-interferon
CA2363779A1 (en) 1999-02-26 2000-08-31 Human Genome Sciences, Inc. Human endokine alpha and methods of use
EP2301947A3 (de) 1999-02-26 2011-11-23 Millennium Pharmaceuticals, Inc. Sekretierte Proteine und Verwendungen dafür
DE19915057A1 (de) 1999-04-01 2000-10-19 Forschungszentrum Borstel Monoklonale Antikörper gegen das humane Mcm3 Protein, Verfahren zu ihrer Herstellung und ihre Verwendung
IL130225A0 (en) 1999-05-31 2000-06-01 Yissum Res Dev Co Novel uses of antibodies against ache and peptides thereof
US20040001826A1 (en) 1999-06-30 2004-01-01 Millennium Pharmaceuticals, Inc. Glycoprotein VI and uses thereof
US7291714B1 (en) 1999-06-30 2007-11-06 Millennium Pharmaceuticals, Inc. Glycoprotein VI and uses thereof
US8557244B1 (en) 1999-08-11 2013-10-15 Biogen Idec Inc. Treatment of aggressive non-Hodgkins lymphoma with anti-CD20 antibody
ES2260059T3 (es) 1999-09-29 2006-11-01 Diagnocure Inc. Rna mensajero del pca3 en tejidos benignos y malignos de prostata.
US7892541B1 (en) 1999-09-30 2011-02-22 Tumor Biology Investment Group, Inc. Soluble epidermal growth factor receptor isoforms
JP2003513660A (ja) 1999-11-05 2003-04-15 バイオジェン インコーポレイテッド ヘッジホッグ融合タンパク質及び利用
US20040002068A1 (en) 2000-03-01 2004-01-01 Corixa Corporation Compositions and methods for the detection, diagnosis and therapy of hematological malignancies
AU2001266557A1 (en) 2000-04-12 2001-10-23 Human Genome Sciences, Inc. Albumin fusion proteins
EP1714661A3 (de) 2000-05-19 2012-03-14 The Center for Blood Research, INC. Verfahren zur Diagnose und Behandlung von hämostatischen Störungen durch Modulation der P-Selectin Aktivität
CA2413160A1 (en) 2000-06-15 2001-12-20 Human Genome Sciences, Inc. Human tumor necrosis factor delta and epsilon
EP2275449B1 (de) 2000-06-16 2016-09-28 Human Genome Sciences, Inc. Immunspezifisch bindende Antikörper gegen BLyS
JP2003000268A (ja) 2000-08-25 2003-01-07 Pfizer Prod Inc 血管新生にかかわる疾患を診断および治療するための方法および組成物
EP1349946B1 (de) 2000-08-25 2011-01-26 BASF Plant Science GmbH Polynukleotide, die für prenylproteasen aus pflanzen kodieren
WO2002046208A2 (en) 2000-11-01 2002-06-13 Elusys Therapeutics, Inc. Method of producing biospecific molecules by protein trans-splicing
AU2001297872B2 (en) 2000-11-17 2006-11-09 University Of Rochester In vitro methods of producing and identifying immunoglobulin molecules in eukaryotic cells
US6989247B2 (en) 2000-11-28 2006-01-24 Celltech R & D, Inc. Compositions and methods for diagnosing or treating psoriasis
PE20020574A1 (es) 2000-12-06 2002-07-02 Wyeth Corp Anticuerpos humanizados que reconocen el peptido amiloideo beta
US7445802B2 (en) 2000-12-26 2008-11-04 Yeda Research And Development Co. Ltd Site-specific in situ generation of allicin using a targeted alliinase delivery system for the treatment of cancers, tumors, infectious diseases and other allicin-sensitive diseases
US7256257B2 (en) 2001-04-30 2007-08-14 Seattle Genetics, Inc. Pentapeptide compounds and uses related thereto
US6972324B2 (en) 2001-05-18 2005-12-06 Boehringer Ingelheim Pharmaceuticals, Inc. Antibodies specific for CD44v6
US7745398B2 (en) 2001-05-31 2010-06-29 Tumor Biology Investment Group, Inc. Soluble ErbB3 and treatment of cancer
US7744882B2 (en) 2001-05-31 2010-06-29 Tumor Biology Investment Group, Inc. Soluble ErbB3 methods of detection and antibodies
US6534059B2 (en) 2001-06-05 2003-03-18 Advanced Biotherapy, Inc. Compositions and methods for treating hyperimmune response in the eye
US6861056B2 (en) 2001-06-05 2005-03-01 Advanced Biotherapy, Inc. Compositions and methods for treating hyperimmune response in the eye
AU2002314495A1 (en) 2001-06-20 2003-01-02 Prochon Biotech Ltd. Antibodies that block receptor protein tyrosine kinase activation, methods of screening for and uses thereof
US7321026B2 (en) 2001-06-27 2008-01-22 Skytech Technology Limited Framework-patched immunoglobulins
US6884619B2 (en) 2001-07-17 2005-04-26 Yale University Inhibition of BEHAB cleavage and primary central nervous system (CNS) tumors
MX339524B (es) 2001-10-11 2016-05-30 Wyeth Corp Composiciones inmunogenicas novedosas para la prevencion y tratamiento de enfermedad meningococica.
DE10151511A1 (de) 2001-10-18 2003-05-08 Basf Lynx Bioscience Ag ee3-Proteinfamilie und zugrundeliegende DNA-Sequenzen
WO2003049673A2 (en) 2001-12-05 2003-06-19 Baylor College Of Medicine Methods and compositions for control of bone formation via modulation of sympathetic tone
EP1458754B1 (de) 2001-12-18 2009-12-09 Endocube SAS Neue mit dem zelltod assoziierte proteine aus der thap familie und par4 verwandte signalwege, die bei der kontrolle von apoptosis involviert sind
US7858297B2 (en) 2001-12-18 2010-12-28 Centre National De La Recherche Scientifique Cnrs Chemokine-binding protein and methods of use
AU2002359721A1 (en) 2001-12-19 2003-07-09 Bristol-Myers Squibb Company Pichia pastoris formate dehydrogenase and uses therefor
WO2003104428A2 (en) 2002-01-21 2003-12-18 Vaccinex, Inc. Gene differentially expressed in breast and bladder cancer and encoded polypeptides
GB0207533D0 (en) 2002-04-02 2002-05-08 Oxford Glycosciences Uk Ltd Protein
US7371383B2 (en) 2002-04-12 2008-05-13 Medimmune, Inc. Recombinant anti-interleukin-9 antibodies
ATE434186T1 (de) 2002-05-07 2009-07-15 Pasteur Institut Screeningverfahren für pp1-wechselwirkende polypeptide oder proteine, peptide, die die bindung von pp1c an die proteine bcl-2, bcl-xl und bcl-w hemmen, und ihre verwendung
US7250551B2 (en) 2002-07-24 2007-07-31 President And Fellows Of Harvard College Transgenic mice expressing inducible human p25
EP2357006B1 (de) 2002-07-31 2015-09-16 Seattle Genetics, Inc. Arzneimittelkonjugate und deren Verwendung zur Behandlung von Krebs, einer Autoimmunerkrankung oder einer Infektionskrankheit
JP2006514823A (ja) 2002-08-20 2006-05-18 ミレニアム ファーマスーティカルズ インク 子宮頸癌の同定、評価、予防および治療を行うための組成物、キットおよび方法
US7785608B2 (en) 2002-08-30 2010-08-31 Wyeth Holdings Corporation Immunogenic compositions for the prevention and treatment of meningococcal disease
AU2003298873B2 (en) 2002-12-06 2011-09-01 Millennium Pharmaceuticals, Inc. Methods for the identification, assessment, and treatment of patients with proteasome inhibition therapy
DK2311848T3 (da) 2002-12-23 2013-10-14 Vical Inc Kodon-optimerede polynukleotidbaserede vacciner mod human cytomegalovirus-infektion
US20050265992A1 (en) 2003-01-03 2005-12-01 The Research Foundation Of State University Of New York F11 receptor (F11R) antagonists as therapeutic agents
US8557957B2 (en) 2003-01-03 2013-10-15 Elizabeth Kornecki Methods of treating disorders by administration of F11 receptor antagonists
JP2007521249A (ja) 2003-01-24 2007-08-02 エラン ファーマシューティカルズ，インコーポレイテッド 脱髄疾患および麻痺のための組成物ならびに再有髄化剤を投与することによるそれらの治療
ES2427853T3 (es) 2003-02-07 2013-11-04 Diagnocure Inc. Procedimiento para detectar cáncer de próstata en una muestra
US20040180387A1 (en) 2003-03-13 2004-09-16 Fujirebio Diagnostics, Inc. Detection of urinary mesothelin-/megakaryocyte potentiating factor-related peptides for assessment of ovarian cancer
US8080642B2 (en) 2003-05-16 2011-12-20 Vical Incorporated Severe acute respiratory syndrome DNA compositions and methods of use
IL158287A0 (en) 2003-10-07 2004-05-12 Yeda Res & Dev Antibodies to nik, their preparation and use
CA2541804A1 (en) 2003-10-07 2005-04-21 Millennium Pharmaceuticals, Inc. Nucleic acid molecules and proteins for the identification, assessment, prevention, and therapy of ovarian cancer
DK2330201T3 (en) * 2003-10-22 2017-07-24 Keck Graduate Inst PROCEDURES FOR SYNTHESIS OF HEATER-MULTIMATE POLYPEPTIDES WHEN USING A HAPLOID COUPLE STRATEGY
PT1725249E (pt) 2003-11-06 2014-04-10 Seattle Genetics Inc Compostos de monometilvalina capazes de conjugação a ligandos
CA2547165C (en) 2003-11-25 2014-07-08 Ira H. Pastan Mutated anti-cd22 antibodies and immunoconjugates
WO2005063992A1 (ja) 2003-12-30 2005-07-14 Suntory Limited 新規血清型のストレプトコッカス ミュータンスおよびその利用
WO2005072341A2 (en) 2004-01-21 2005-08-11 Fujirebio America, Inc. Detection of mesothelin-/megakaryocyte potentiating factor-related peptides in peritoneal fluid for assessment of the peritoneum and the peritoneal cavity
DK1734996T3 (da) 2004-04-02 2013-06-10 Univ California Fremgangsmåder og sammensætninger til behandling og forebyggelse af sygdom, der er associeret med alfa v beta 5-integrin
CA2566355C (en) 2004-05-18 2014-04-15 Vical Incorporated Influenza virus vaccine composition and methods of use
EP1598428A1 (de) 2004-05-18 2005-11-23 Georg Dewald Verfahren und Kits zum Nachweis von erblichen angioedema typ III
GB0414054D0 (en) 2004-06-23 2004-07-28 Owen Mumford Ltd Improvements relating to automatic injection devices
WO2006012451A2 (en) 2004-07-22 2006-02-02 Five Prime Therapeutics, Inc. Compositions and methods of use for mgd-csf in disease treatment
CA2582157A1 (en) 2004-10-05 2006-04-20 Wyeth Methods and compositions for improving recombinant protein production
US7462454B2 (en) 2004-10-12 2008-12-09 Advanced Biotherapy, Inc. Treatment of herpes
JP2008519863A (ja) 2004-11-12 2008-06-12 シアトル ジェネティクス インコーポレイティッド Ｎ末端にアミノ安息香酸単位を有するオーリスタチン
EP1838348B1 (de) 2004-12-15 2013-06-26 Janssen Alzheimer Immunotherapy Humanisierte amyloid-beta-antikörper zur verbesserung der kognition
CA2491067A1 (en) 2004-12-24 2006-06-24 Stichting Katholieke Universiteit Mrna rations in urinary sediments and/or urine as a prognostic marker for prostate cancer
AU2006214384A1 (en) 2005-02-14 2006-08-24 University Of Pittsburgh Of The Commonwealth System Of Higher Education Use of IL-17F in diagnosis and therapy of airway inflammation
SI2567976T1 (sl) 2005-03-23 2017-11-30 Genmab A/S Protitelesa usmerjena proti cd38 za zdravljenje multiplega mieloma
EP2343320B1 (de) 2005-03-25 2017-10-25 GITR, Inc. Anti-gitr antikörper und ihre verwendung
AU2006236225C1 (en) 2005-04-19 2013-05-02 Seagen Inc. Humanized anti-CD70 binding agents and uses thereof
ES2547463T3 (es) 2005-06-17 2015-10-06 Merck Sharp & Dohme Corp. Moléculas de unión a ILT3 y usos de las mismas
CA2611816C (en) 2005-06-17 2014-05-27 Wyeth Methods of purifying fc region containing proteins
ES2708763T3 (es) 2005-07-07 2019-04-11 Seattle Genetics Inc Compuestos de monometilvalina que tienen modificaciones de la cadena lateral de fenilalanina en el extremo C
US8039273B2 (en) 2005-07-18 2011-10-18 Seattle Genetics, Inc. β-glucuronide-linker drug conjugates
NZ589687A (en) 2005-07-21 2012-09-28 Genmab As Potency assays for antibody drug substance binding to an FC receptor
WO2007024940A2 (en) 2005-08-22 2007-03-01 Dana-Farber Cancer Institute, Inc. Mitochondrial localization of muc1
ATE547709T1 (de) 2005-11-14 2012-03-15 Metamol Theranostics Llc Tumorinvasionsfördernde peptidsequenz
US10183986B2 (en) 2005-12-15 2019-01-22 Industrial Technology Research Institute Trimeric collagen scaffold antibodies
EP1806365A1 (de) 2006-01-05 2007-07-11 Boehringer Ingelheim International GmbH Antikörper spezifisch für Fibroblasten-Aktivierungsprotein und Immunokonjugaten, die diese Antikörper enthalten
CA2647671A1 (en) 2006-03-31 2007-11-01 Massachusetts Institute Of Technology Treatment of tumors expressing mutant egf receptors
CA2646319A1 (en) 2006-04-20 2007-11-01 The Henry M. Jackson Foundation For The Advancement Of Military Medicine , Incorporated Methods and compositions based on shiga toxin type 1 protein
GB0611116D0 (en) 2006-06-06 2006-07-19 Oxford Genome Sciences Uk Ltd Proteins
CA2655903A1 (en) 2006-06-19 2008-08-07 Tolerx, Inc. Ilt3 binding molecules and uses therefor
AU2007264672B2 (en) 2006-06-28 2013-03-14 Hadasit Medical Research Services & Development Ltd Caspase-8 and inflammation, infection and wound healing
CN101484199B (zh) 2006-06-30 2014-06-25 艾伯维生物技术有限公司 自动注射装置
US7572618B2 (en) 2006-06-30 2009-08-11 Bristol-Myers Squibb Company Polynucleotides encoding novel PCSK9 variants
JP5605895B2 (ja) 2006-07-04 2014-10-15 ゲンマブ エー／エス Ｃｏｐｄを処置するためのｃｄ２０結合分子
US20090203602A1 (en) 2006-09-01 2009-08-13 Cohava Gelber Compositions and methods for diagnosis and treatment of type 2 diabetes
US7951776B2 (en) 2006-09-01 2011-05-31 American Type Culture Collection Methods for treatment of type 1 diabetes
ME03503B (de) 2006-09-26 2020-04-20 Genmab As Anti-cd38 plus kortikosteroids plus ein nicht kortikosteroid chemotherapeutikum zur behandlung von tumoren
EP1916259A1 (de) 2006-10-26 2008-04-30 Institut National De La Sante Et De La Recherche Medicale (Inserm) Anti-Glycoprotein VI SCFV Fragment zur Thrombosebehandlung
CA2669095A1 (en) 2006-11-15 2008-05-29 Functional Genetics, Inc. Anti-tsg101 antibodies and their uses for treatment of viral infections
CA2699837C (en) 2006-12-01 2017-06-13 Seattle Genetics, Inc. Variant target binding agents and uses thereof
HUE041957T2 (hu) 2006-12-01 2019-06-28 Novartis Ag Anti-P-szelektin antitestek és eljárások azok alkalmazására gyulladásos betegségek kezelésére
AR064642A1 (es) 2006-12-22 2009-04-15 Wyeth Corp Polinucleotido vector que lo comprende celula recombinante que comprende el vector polipeptido , anticuerpo , composicion que comprende el polinucleotido , vector , celula recombinante polipeptido o anticuerpo , uso de la composicion y metodo para preparar la composicion misma y preparar una composi
EP2115173B1 (de) 2007-01-08 2011-09-21 Government of the USA, as Represented by the Secretary, Department of Health and Human Services Slco1b3-genotyp
ES2426158T3 (es) 2007-01-22 2013-10-21 Genentech, Inc. Precipitación con polielectrolito y purificación de anticuerpos
WO2008101121A2 (en) 2007-02-14 2008-08-21 Dana-Farber Cancer Institute, Inc. Methods and compositions relating to promoter regulation by muc1 and klf proteins
EP2121745A2 (de) 2007-02-26 2009-11-25 Oxford Genome Sciences (UK) Limited Proteine
WO2008104803A2 (en) 2007-02-26 2008-09-04 Oxford Genome Sciences (Uk) Limited Proteins
WO2008106785A1 (en) 2007-03-05 2008-09-12 Cancer Care Ontario Assessment of risk for colorectal cancer
US9707299B2 (en) 2007-05-23 2017-07-18 The Trustees Of The University Of Pennsylvania Targeted carriers for intracellular drug delivery
EP2666787B1 (de) 2007-05-31 2022-02-09 Genmab A/S STABILE IgG4-ANTIKÖRPER
US8591886B2 (en) 2007-07-12 2013-11-26 Gitr, Inc. Combination therapies employing GITR binding molecules
CA2695505A1 (en) 2007-08-06 2009-02-12 Burnham Institute For Medical Research Znf206: a novel regulator of embryonic stem cell self-renewal and pluripotency
PE20140196A1 (es) 2007-08-09 2014-03-19 Boehringer Ingelheim Int Anticuerpos anti-cd37
MX2010003220A (es) 2007-09-24 2010-08-04 Univ Cornell Proteinas inmunogenicas de membrana exterior derivada de genoma de leptospira y composiciones y metodos basados en las mismas.
NZ584642A (en) 2007-09-27 2012-08-31 Japan Tobacco Inc Factor involved in latent infection with herpesvirus, and use thereof
GB0719231D0 (en) 2007-10-03 2007-11-14 Oxford Genome Sciences Uk Ltd Protein
JO3076B1 (ar) 2007-10-17 2017-03-15 Janssen Alzheimer Immunotherap نظم العلاج المناعي المعتمد على حالة apoe
JP6018361B2 (ja) 2008-01-31 2016-11-02 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル 制御性ｔ細胞活性を抑制するための、ヒトｃｄ３９に対する抗体およびその使用
DK2265283T3 (da) 2008-03-18 2014-10-20 Seattle Genetics Inc Auristatin-lægemiddel-linker-konjugater
PL2282773T3 (pl) 2008-05-02 2014-08-29 Seattle Genetics Inc Sposoby i kompozycje do wytwarzania przeciwciał i pochodnych przeciwciał o obniżonej fukozylacji rdzeniowej
AU2009269099B2 (en) 2008-07-09 2016-03-10 Biogen Ma Inc. Compositions comprising antibodies to LINGO or fragments thereof
KR101924874B1 (ko) 2008-09-26 2018-12-04 다나-파버 캔서 인스티튜트 인크. 인간 항-ｐｄ-１, ｐｄ-ｌ１, 및 ｐｄ-ｌ2 항체 및 그의 용도
JP2012504946A (ja) 2008-10-07 2012-03-01 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル 血小板第４因子変異体１（ｐｆ４ｖ１）に対する中和抗体およびそのフラグメント
US9067981B1 (en) 2008-10-30 2015-06-30 Janssen Sciences Ireland Uc Hybrid amyloid-beta antibodies
CN102292101A (zh) 2008-11-26 2011-12-21 五佳医疗股份有限公司 通过serpine2来调节胶原和平滑肌肌动蛋白表达的组合物和方法
CN102325790A (zh) 2008-12-03 2012-01-18 西马生物医学计划公司 应用酚可溶性调控蛋白研制疫苗
WO2010084408A2 (en) 2009-01-21 2010-07-29 Oxford Biotherapeutics Ltd. Pta089 protein
WO2010088522A2 (en) 2009-01-30 2010-08-05 Ab Biosciences, Inc. Novel lowered affinity antibodies and uses therefor
WO2010106051A1 (en) 2009-03-17 2010-09-23 Universite De La Mediterranee Btla antibodies and uses thereof
EP2427497B1 (de) 2009-05-07 2016-12-07 Stallergenes Verwendung von igg1-immunoglobulinen und/oder liganden des cd32-rezeptors zur behandlung entzündlicher krankheiten und ereignisse über die schleimhaut
EP2272979A1 (de) 2009-06-30 2011-01-12 Centre National de la Recherche Scientifique (CNRS) Verfahren zum Testen einer Person, die eine Prädisposition für Krebs aufzuweisen scheint
IN2012DN00303A (de) 2009-07-24 2015-05-08 Univ California
JP5795765B2 (ja) 2009-09-11 2015-10-14 アメリカ合衆国 低減した免疫原性を有する改良型シュードモナス（Ｐｓｅｕｄｏｍｏｎａｓ）外毒素Ａ
US9547007B2 (en) 2009-09-22 2017-01-17 The Trustees Of The University Of Pennslyvania Methods and compositions for in-vivo enzyme capture
US20120263719A1 (en) 2009-10-22 2012-10-18 Yeda Research And Development Co., Ltd. Compositions and methods for treating aspergillosis
EP2496605A1 (de) 2009-11-02 2012-09-12 Oxford Biotherapeutics Ltd. Ror1 als therapeutisches und diagnostisches ziel
WO2011080322A1 (en) 2009-12-30 2011-07-07 Institut National De La Sante Et De La Recherche Medicale (Inserm) Method of prognosing the outcome of acquired hemophilia and of treatment of hemophilia
HUE033044T2 (hu) 2010-01-26 2017-11-28 Nat Jewish Health Eljárás tüdõ rendellenességek kockázatának elõrejelzésére, diagnózisára, kórjóslatára
JP6101489B2 (ja) 2010-01-28 2017-03-22 アブ バイオサイエンシズ インコーポレイテッド 親和性が低下した抗体およびそれを作製する方法
TWI518325B (zh) 2010-02-04 2016-01-21 自治醫科大學 對ａｌｋ抑制劑具有先抗性或後抗性癌症的識別、判斷和治療
GB201003701D0 (en) * 2010-03-05 2010-04-21 Cilian Ag System for the expression of a protein
US10745467B2 (en) 2010-03-26 2020-08-18 The Trustees Of Dartmouth College VISTA-Ig for treatment of autoimmune, allergic and inflammatory disorders
US20150231215A1 (en) 2012-06-22 2015-08-20 Randolph J. Noelle VISTA Antagonist and Methods of Use
CA2794483C (en) 2010-03-26 2020-07-07 Trustees Of Dartmouth College Vista regulatory t cell mediator protein, vista binding agents and use thereof
US20120100166A1 (en) 2010-07-15 2012-04-26 Zyngenia, Inc. Ang-2 Binding Complexes and Uses Thereof
EP2596025B1 (de) 2010-07-23 2018-06-20 INSERM (Institut National de la Santé et de la Recherche Médicale) Gegen co-029 gerichtete krebstherapieverfahren
EP2412724A1 (de) 2010-07-29 2012-02-01 Centre National de la Recherche Scientifique (C.N.R.S) Regulierung der Glypican-4-Aktivität zur Modulierung des Schicksals von Stammzellen und Verwendungen davon
ES2639035T3 (es) 2010-08-23 2017-10-25 Wyeth Llc Formulaciones estables de antígenos rLP2086 de Neisseria meningitidis
ES2585328T5 (es) 2010-09-10 2022-12-14 Wyeth Llc Variantes no lipidadas de antígenos ORF2086 de Neisseria meningitidis
UA112062C2 (uk) 2010-10-04 2016-07-25 Бьорінгер Інгельхайм Інтернаціональ Гмбх Cd33-зв'язувальний агент
WO2012064743A2 (en) 2010-11-08 2012-05-18 The Johns Hopkins University Methods for improving heart function
WO2012071513A2 (en) 2010-11-24 2012-05-31 Hong Gao Expanding hematopoietic stem cells
EP2648754A4 (de) 2010-12-07 2016-02-24 Philadelphia Health & Educatio Verfahren zur hemmung von krebs-metastasen
WO2012080769A1 (en) 2010-12-15 2012-06-21 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-cd277 antibodies and uses thereof
DK2654789T3 (en) 2010-12-22 2018-09-03 Orega Biotech ANTIBODIES AGAINST HUMAN CD39 AND USE THEREOF
US20150018408A1 (en) 2013-07-10 2015-01-15 The Regents Of The University Of Michigan Therapeutic antibodies and uses thereof
WO2012096960A2 (en) 2011-01-10 2012-07-19 The Regents Of The University Of Michigan Stem cell factor inhibitor
EP3187216B1 (de) 2011-01-24 2019-08-21 AbbVie Biotechnology Ltd. Automatische injektionsvorrichtungen mit umspritzten greifflächen
WO2012101125A1 (en) 2011-01-24 2012-08-02 INSERM (Institut National de la Santé et de la Recherche Médicale) Specific antibodies against human cxcl4 and uses thereof
CA2828972C (en) 2011-03-02 2021-09-07 Berg Llc Interrogatory cell-based assays and uses thereof
CN103517922B (zh) 2011-03-31 2016-10-19 国家医疗保健研究所 抗icos的抗体及其用途
US20120328567A1 (en) 2011-04-08 2012-12-27 Steven Bushnell Biomarkers predictive of therapeutic responsiveness to ifnb and uses thereof
US10654916B2 (en) 2011-04-21 2020-05-19 The Regents Of The University Of California, A California Corporation Compositions and methods for the treatment of neuromyelitis optica
CN103619378B (zh) 2011-04-21 2017-03-01 艾伯维公司 可佩戴式自动注射装置
SG194787A1 (en) 2011-05-06 2013-12-30 Us Gov Health & Human Serv Recombinant immunotoxin targeting mesothelin
CN103890010B (zh) 2011-05-19 2017-04-19 法国国家健康医学研究院 抗‑人‑her3抗体及其用途
CN106432506A (zh) 2011-05-24 2017-02-22 泽恩格尼亚股份有限公司 多价和单价多特异性复合物及其用途
JP6100764B2 (ja) 2011-06-09 2017-03-22 アメリカ合衆国 免疫原性の低いｔ細胞及び／又はｂ細胞エピトープを有するシュードモナス外毒素ａ
AU2012273954A1 (en) 2011-06-22 2014-01-09 Inserm (Institut National De La Sante Et De La Recherche Medicale) Anti-Axl antibodies and uses thereof
RU2014101707A (ru) 2011-06-22 2015-07-27 Инсерм (Энститю Насьональ Де Ля Сантэ Э Де Ля Решерш Медикаль) АНТИ-Axl АНТИТЕЛА И ИХ ПРИМЕНЕНИЕ
EP2726094B1 (de) 2011-06-28 2016-12-14 Oxford BioTherapeutics Ltd Therapeutisches und diagnostisches ziel
EP2543679A1 (de) 2011-07-08 2013-01-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Antikörper zur Behandlung und Vorbeugung von Thrombose
EP2543677A1 (de) 2011-07-08 2013-01-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Antikörper zur Behandlung und Vorbeugung von Thrombose
EP2543678A1 (de) 2011-07-08 2013-01-09 INSERM (Institut National de la Santé et de la Recherche Médicale) Antikörper zur Behandlung und Vorbeugung von Thrombose
DE202011103324U1 (de) 2011-07-12 2012-01-02 Nekonal S.A.R.L. Therapeutische anti-TIRC7 Antikörper für die Verwendung in Immun und anderen Krankheiten
US20150125533A1 (en) 2011-07-25 2015-05-07 American University In Cairo Single-domain antibodies and graphene coated magnetic metal nanoparticles conjugate and methods for using the same
BR112014004162A2 (pt) 2011-08-22 2018-04-24 Cangene Corp anticorpos contra clostridium difficile
ES2704126T3 (es) 2011-08-23 2019-03-14 Found Medicine Inc Moléculas de fusión KIF5B-RET y usos de las mismas
CA2846667A1 (en) 2011-09-06 2013-03-14 Guy L. Reed Serpinf2-binding molecules and methods of use
EP3301110A1 (de) 2011-09-16 2018-04-04 The USA, as represented by The Secretary, Department of Health and Human Services Pseudomonas exotoxin a mit weniger immunogenen b-zellepitopen
WO2013056178A2 (en) 2011-10-14 2013-04-18 Foundation Medicine, Inc. Novel estrogen receptor mutations and uses thereof
PT2780039T (pt) 2011-11-17 2018-01-29 Pfizer Péptidos citotóxicos e seus conjugados de anticorpos e fármacos
CN105693830B (zh) 2012-03-09 2021-07-06 辉瑞公司 脑膜炎双球菌组合物及其方法
SA115360586B1 (ar) 2012-03-09 2017-04-12 فايزر انك تركيبات لعلاج الالتهاب السحائي البكتيري وطرق لتحضيرها
AR090340A1 (es) 2012-03-27 2014-11-05 Genentech Inc Operaciones de cosecha mejoradas para proteinas recombinadas
US9494597B2 (en) 2012-04-02 2016-11-15 Ab Biosciences, Inc. Human control antibodies and uses therefor
CA2869296A1 (en) 2012-04-02 2013-10-10 Berg Llc Interrogatory cell-based assays and uses thereof
EA030716B1 (ru) 2012-05-14 2018-09-28 Байоджен Ма Инк. Антагонисты lingo-2 для лечения заболеваний, в которых участвуют двигательные нейроны
ES2983109T3 (es) 2012-05-15 2024-10-21 Seagen Inc Conjugados enlazadores autoestabilizantes
US9890215B2 (en) 2012-06-22 2018-02-13 King's College London Vista modulators for diagnosis and treatment of cancer
US9676847B2 (en) 2012-06-25 2017-06-13 Orega Biotech IL-17 antagonist antibodies
US20150175979A1 (en) 2012-07-23 2015-06-25 La Jolla Institute For Allergy And Immunology Ptprs and proteoglycans in autoimmune disease
GB201213652D0 (en) 2012-08-01 2012-09-12 Oxford Biotherapeutics Ltd Therapeutic and diagnostic target
WO2014033327A1 (en) 2012-09-03 2014-03-06 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies directed against icos for treating graft-versus-host disease
JP6368308B2 (ja) 2012-09-07 2018-08-01 トラスティーズ・オブ・ダートマス・カレッジ 癌の診断および治療のためのｖｉｓｔａ調節剤
AU2013337277B2 (en) 2012-11-05 2018-03-08 Foundation Medicine, Inc. Novel NTRK1 fusion molecules and uses thereof
PL2917195T3 (pl) 2012-11-05 2018-04-30 Pfizer Inc. Analogi spliceostatyny
KR20150082548A (ko) 2012-11-07 2015-07-15 화이자 인코포레이티드 항-notch3 항체 및 항체-약물 접합체
UY35147A (es) 2012-11-20 2014-06-30 Sanofi Sa Anticuerpos anti-ceacam5 y usos de éstos.
WO2014089177A2 (en) 2012-12-04 2014-06-12 Massachusetts Institute Of Technology Compounds, conjugates and compositions of epipolythiodiketopiperazines and polythiodiketopiperazines
JP6441232B2 (ja) 2012-12-27 2018-12-19 サノフイＳａｎｏｆｉ 抗ｌａｍｐ１抗体および抗体薬物コンジュゲート、ならびにその使用
CA3150658A1 (en) 2013-01-18 2014-07-24 Foundation Medicine, Inc. Methods of treating cholangiocarcinoma
GB201302447D0 (en) 2013-02-12 2013-03-27 Oxford Biotherapeutics Ltd Therapeutic and diagnostic target
WO2014130922A1 (en) 2013-02-25 2014-08-28 Trustees Of Boston University Compositions and methods for treating fungal infections
EP2964665B1 (de) 2013-03-08 2018-08-01 Pfizer Inc Immunogene fusionspolypeptide
US10150800B2 (en) 2013-03-15 2018-12-11 Zyngenia, Inc. EGFR-binding modular recognition domains
TR201911139T4 (tr) 2013-03-15 2019-08-21 Zymeworks Inc Sitotoksik ve anti-mitotik bileşikler ve bunları kullanmanın yöntemleri.
US9969804B2 (en) 2013-04-01 2018-05-15 The Regents Of The University Of California Methods and compositions for treating and preventing disease associated with alpha-V beta-8 integrin
US10005839B2 (en) 2013-05-17 2018-06-26 Inserm (Institut National De La Sante Et De La Recherche Medicale) Antagonist of the BTLA/HVEM interaction for use in therapy
US9428748B2 (en) 2013-06-17 2016-08-30 Hong Gao Method of expanding hematopoietic stem cells
WO2014205187A1 (en) 2013-06-20 2014-12-24 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Cytolethal distending toxin subunit b conjugated or fused to bacillus anthracis toxin lethal factor
WO2015003114A1 (en) 2013-07-05 2015-01-08 University Of Washington Through Its Center For Commercialization Soluble mic neutralizing monoclonal antibody for treating cancer
CN105658230B (zh) 2013-08-29 2020-04-21 希望之城 细胞穿透缀合物及其使用方法
WO2015033251A2 (en) 2013-09-08 2015-03-12 Pfizer Inc. Neisseria meningitidis compositions and methods thereof
CA2926215A1 (en) 2013-10-06 2015-04-09 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Modified pseudomonas exotoxin a
NZ758050A (en) 2013-10-15 2024-03-22 Seagen Inc Pegylated drug-linkers for improved ligand-drug conjugate pharmacokinetics
EP3066124B1 (de) 2013-11-07 2021-01-06 INSERM - Institut National de la Santé et de la Recherche Médicale Gegen her3, neuregulin allosterische antikörpern
JP6667445B2 (ja) 2013-11-15 2020-03-18 アンスティテュ・パストゥール 熱帯熱マラリア原虫アルテミシニン耐性の分子マーカー
WO2015091653A2 (en) 2013-12-17 2015-06-25 Westfaelische Wilhelms-Universitaet Muenster Means and methods for treating a pruritus-like skin-disease
DK3082878T3 (da) 2013-12-19 2022-12-12 Seagen Inc Methylencarbamatlinkere til anvendelse med targeterede lægemiddelkonjugater
WO2015095868A1 (en) 2013-12-20 2015-06-25 Wake Forest University Health Sciences Methods and compositions for increasing protective antibody levels induced by pneumococcal polysaccharide vaccines
EP2960252A1 (de) 2014-06-26 2015-12-30 Institut Pasteur Phospholipase zur Behandlung von Immunosuppression
US11014987B2 (en) 2013-12-24 2021-05-25 Janssen Pharmaceutics Nv Anti-vista antibodies and fragments, uses thereof, and methods of identifying same
EP3712174B1 (de) 2013-12-24 2022-03-16 Janssen Pharmaceutica NV Anti-vista-antikörper und -fragmente
RU2729194C2 (ru) 2013-12-27 2020-08-05 Займворкс Инк. Сульфонамидсодержащие связывающие системы для лекарственных конъюгатов
WO2015103026A2 (en) 2014-01-03 2015-07-09 The Regents Of The University Of Michigan Treatment of neurological disorders
EP3099692B9 (de) 2014-01-27 2019-11-13 Pfizer Inc Bifunktionelle zytostatika
KR102647074B1 (ko) 2014-02-17 2024-03-14 씨젠 인크. 친수성 항체-약물 컨쥬게이트
EP3145546B1 (de) 2014-02-19 2020-12-09 Emergent BioSolutions Canada Inc. Monoklonale marburg-antikörper
TWI609886B (zh) 2014-04-10 2018-01-01 台灣浩鼎生技股份有限公司 抗體、產生該抗體之融合瘤、包含該抗體之藥學組成物及其用途
US10544231B2 (en) 2014-04-16 2020-01-28 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies for the prevention or the treatment of bleeding episodes
CA2946606C (en) 2014-05-13 2023-06-06 Bavarian Nordic A/S Combination therapy for treating cancer with a poxvirus expressing a tumor antigen and an antagonist of tim-3
CN107073109B (zh) 2014-06-11 2021-08-06 凯西·A·格林 Vista激动剂和拮抗剂抑制或增强体液免疫的用途
US10414814B2 (en) 2014-07-03 2019-09-17 City Of Hope Tumor-selective CTLA-4 antagonists
JP6789124B2 (ja) 2014-07-03 2020-11-25 イエール ユニバーシティ 腫瘍形成を抑制するディコップ２（Ｄｉｃｋｋｏｐｆ２）（Ｄｋｋ２）阻害
US10752668B2 (en) 2014-07-25 2020-08-25 Theravectys Lentiviral vectors for regulated expression of a chimeric antigen receptor molecule
US10392444B2 (en) 2014-08-08 2019-08-27 Oncoquest, Inc. Tumor antigen specific antibodies and TLR3 stimulation to enhance the performance of checkpoint interference therapy of cancer
US10278986B2 (en) 2014-08-14 2019-05-07 The Regents Of The University Of Colorado, A Body Corporate Antibody-siRNA conjugates and uses therefor
WO2016044234A1 (en) 2014-09-16 2016-03-24 Eric Tsao Anti-egfr antibody and uses of same
SG11201702143PA (en) 2014-09-17 2017-04-27 Zymeworks Inc Cytotoxic and anti-mitotic compounds, and methods of using the same
JP2017531642A (ja) 2014-10-03 2017-10-26 マサチューセッツ インスティテュート オブ テクノロジー エボラ糖タンパク質に結合する抗体およびその使用
MX2017007136A (es) 2014-12-05 2017-12-04 Immunext Inc Identificacion de vsig8 como el receptor vista putativo y su uso para producir moduladores vista/vsig8.
MX2017009038A (es) 2015-01-08 2017-10-25 Biogen Ma Inc Antagonistas de proteina 1 de interacción con el receptor de nogo que contiene el dominio de inmunoglobulina y repeticiones ricas en leucina (lingo-1) y usos para el tratamiento de trastornos desmielinizantes.
EP3733696A1 (de) 2015-01-13 2020-11-04 City of Hope Peptid-linkermasken mit ctla4-bindenden proteinen
EP3244924B1 (de) 2015-01-15 2021-04-07 Oncoquest Pharmaceuticals Inc. Verfahren zur steigerung der abgabe von anti-krebsmitteln an ziele
SG11201705413WA (en) 2015-01-16 2017-08-30 Hope City Cell penetrating antibodies
CN107428835B (zh) 2015-01-23 2021-11-26 赛诺菲 抗cd3抗体、抗cd123抗体和与cd3和/或cd123特异性结合的双特异性抗体
WO2016128349A1 (en) 2015-02-09 2016-08-18 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies specific to glycoprotein (gp) of ebolavirus and uses for the treatment and diagnosis of ebola virus infection
BR112017017460A2 (pt) 2015-02-19 2018-04-10 Pfizer Inc. composições de neisseria meningitidis e métodos das mesmas
KR102046862B1 (ko) 2015-03-20 2019-11-20 화이자 인코포레이티드 Cti 약물작용발생단을 함유하는 이관능성 세포독성제
CA2981504C (en) 2015-03-30 2023-10-24 City Of Hope Mechanically interlocking complexes
US20180071413A1 (en) 2015-04-07 2018-03-15 Inserm (Institut National De La Sante Et De La Recherche Medicale) Non-invasive imaging of tumor pd-l1 expression
EP3285811A1 (de) 2015-04-21 2018-02-28 Institut Gustave Roussy Therapeutische verfahren, produkte und zusammensetzungen zur hemmung von znf555
US9574014B2 (en) 2015-05-15 2017-02-21 City Of Hope Chimeric antigen receptor compositions
US11009509B2 (en) 2015-06-24 2021-05-18 Janssen Pharmaceutica Nv Anti-VISTA antibodies and fragments
JOP20200312A1 (ar) 2015-06-26 2017-06-16 Novartis Ag الأجسام المضادة للعامل xi وطرق الاستخدام
US20180201687A1 (en) 2015-07-07 2018-07-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity to myosin 18a and uses thereof
EP3331543A4 (de) 2015-08-06 2019-03-20 City of Hope Zellpenetrierende proteinantikörperkonjugate und verfahren zur verwendung
WO2017031353A1 (en) 2015-08-19 2017-02-23 Rutgers, The State University Of New Jersey Novel methods of generating antibodies
AU2016315892B2 (en) 2015-09-02 2023-06-15 The Regents Of The University Of Colorado, A Body Corporate Compositions and methods for modulating T-cell mediated immune response
MX2018002708A (es) 2015-09-04 2018-08-01 Primatope Therapeutics Inc Anticuerpos anti-cd40 humanizados y usos de los mismos.
US10501532B2 (en) 2015-10-07 2019-12-10 Obi Pharma, Inc. Carbohydrate antibodies, pharmaceutical compositions and uses thereof
WO2017060397A1 (en) 2015-10-09 2017-04-13 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods for predicting the survival time of subjects suffering from melanoma metastases
CA3003252C (en) 2015-10-28 2024-06-25 Yale University Humanized anti-dkk2 antibody and uses thereof
CN107043406B (zh) 2015-11-03 2021-08-17 财团法人工业技术研究院 化合物、连接子-药物、及配体-药物耦合体
WO2017091683A1 (en) 2015-11-25 2017-06-01 Visterra, Inc. Antibody molecules to april and uses thereof
AU2016363013B2 (en) 2015-12-04 2022-03-10 Seagen Inc. Conjugates of quaternized tubulysin compounds
US11793880B2 (en) 2015-12-04 2023-10-24 Seagen Inc. Conjugates of quaternized tubulysin compounds
CN112274649A (zh) 2015-12-10 2021-01-29 希望之城 细胞穿透花青偶联抗体
EP3389713A2 (de) 2015-12-17 2018-10-24 Novartis AG Kombination von c-met-inhibitor mit antikörpermolekül zu pd-1 und verwendungen davon
WO2017106806A1 (en) 2015-12-18 2017-06-22 Federica Cavallo COMPOSITIONS AND METHODS RELATED TO xCT PEPTIDES
EA201891509A1 (ru) 2015-12-24 2019-01-31 Корвус Фармасьютикалз, Инк. Способы лечения рака
WO2017134547A1 (en) 2016-02-01 2017-08-10 Pfizer Inc. Tubulysin analogs and methods for their preparation
EP3410849B1 (de) 2016-02-05 2023-07-05 Institut Pasteur Verwendung von inhibitoren von adam12 als adjuvanzien in tumortherapien
MX2018009800A (es) 2016-02-12 2018-11-09 Janssen Pharmaceutica Nv Anticuerpos y fragmentos anti-vista, usos de los mismos y procedimientos de identificacion de los mismos.
WO2017144668A1 (en) 2016-02-26 2017-08-31 INSERM (Institut National de la Santé et de la Recherche Médicale) Antibodies having specificity for btla and uses thereof
AU2017225869A1 (en) 2016-02-29 2018-09-06 Foundation Medicine, Inc. Methods of treating cancer
CA3017356A1 (en) 2016-03-10 2017-09-14 Aperisys, Inc. Antigen-binding fusion proteins with modified hsp70 domains
BR112018069273A2 (pt) 2016-03-25 2019-01-22 Seattle Genetics Inc métodos para preparação de um composto e para tratamento de um indivíduo com uma malignidade hematológica, composto, composição, e, intermediário ligante de fármaco ou composto ligante de fármaco
AU2017250294B2 (en) 2016-04-15 2022-07-21 Immunext Inc. Anti-human VISTA antibodies and use thereof
WO2017197045A1 (en) 2016-05-11 2017-11-16 Movassaghi Mohammad Convergent and enantioselective total synthesis of communesin analogs
TW201802121A (zh) 2016-05-25 2018-01-16 諾華公司 抗因子XI/XIa抗體之逆轉結合劑及其用途
US11780924B2 (en) 2016-06-21 2023-10-10 University Of Oslo HLA binding vaccine moieties and uses thereof
JP7219207B2 (ja) 2016-07-29 2023-02-07 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル 腫瘍関連マクロファージを標的化する抗体及びその使用
WO2018025168A1 (en) 2016-08-03 2018-02-08 Pfizer Inc. Heteroaryl sulfone-based conjugation handles, methods for their preparation, and their use in synthesizing antibody drug conjugates
MX2019001302A (es) 2016-08-09 2019-06-12 Seattle Genetics Inc Conjugados de farmaco con enlazadores autoestabilizantes que tienen propiedades fisioquimicas mejoradas.
ES2919927T3 (es) 2016-08-10 2022-07-29 Pasteur Institut Métodos y reactivos para la detección del paludismo por Plasmodium falciparum resistente a la piperaquina
RS60807B1 (sr) 2016-08-15 2020-10-30 Novartis Ag Režimi i postupci za lečenje multiple skleroze upotrebom ofatumumaba
JP7274413B2 (ja) 2016-09-23 2023-05-16 マレンゴ・セラピューティクス，インコーポレーテッド ラムダ及びカッパ軽鎖を含む多重特異性抗体分子
MX2019004371A (es) 2016-10-13 2019-11-18 Massachusetts Inst Technology Anticuerpos que se unen a la proteina de envoltura del virus zika y usos de los mismos.
TWI835714B (zh) 2016-10-18 2024-03-21 美商思進公司 菸鹼醯胺腺嘌呤二核苷酸補救合成(salvage pathway)抑制劑之靶向投遞
US11286295B2 (en) 2016-10-20 2022-03-29 Sanofi Anti-CHIKV monoclonal antibodies directed against the E2 structural protein
EP3538101B1 (de) 2016-11-14 2024-06-19 Virginia Commonwealth University Inhibitoren von krebs und/oder metastasis
JP7244987B2 (ja) 2016-12-14 2023-03-23 シージェン インコーポレイテッド 多剤抗体薬物コンジュゲート
WO2018119380A2 (en) 2016-12-23 2018-06-28 Bristol-Myers Squibb Company Design of therapeutic immunoglobulin g4 for improved bioanalytical and bioprocessing properties
US11168147B2 (en) 2016-12-23 2021-11-09 Novartis Ag Factor XI antibodies and methods of use
US11993625B2 (en) 2017-01-24 2024-05-28 Pfizer, Inc. Calicheamicin derivatives and antibody drug conjugates thereof
US10183070B2 (en) 2017-01-31 2019-01-22 Pfizer Inc. Neisseria meningitidis compositions and methods thereof
US20180221476A1 (en) 2017-02-06 2018-08-09 Oncoquest Nc. Treatment of cancer with therapeutic monoclonal antibody specific for a tumor associated antigen and an immune adjuvant
JP7116736B2 (ja) 2017-03-02 2022-08-10 ノバルティス アーゲー 操作されたヘテロ二量体タンパク質
JP2020510432A (ja) 2017-03-02 2020-04-09 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル Ｎｅｃｔｉｎ−４への特異性を有する抗体及びその使用
CA3051640A1 (en) 2017-03-16 2018-09-20 Innate Pharma Compositions and methods for treating cancer
WO2018176019A1 (en) 2017-03-24 2018-09-27 The Regents Of The University Of California Proteoglycan irregularities in abnormal fibroblasts and therapies based therefrom
CN110430901B (zh) 2017-03-24 2024-08-16 西雅图基因公司 制备葡糖苷酸药物-接头及其中间体的方法
MX2019011858A (es) 2017-04-03 2019-11-21 Oncologie Inc Metodos para tratar el cancer con anticuerpos dirigidos a ps con agentes inmunooncologicos.
JP2020517256A (ja) 2017-04-19 2020-06-18 エルスター セラピューティクス， インコーポレイテッド 多重特異性分子およびその使用
AU2018254776B2 (en) 2017-04-22 2022-06-30 Immunomic Therapeutics, Inc. Improved LAMP constructs
MX2019012194A (es) 2017-04-27 2019-12-16 Seattle Genetics Inc Conjugados inhibidores de la via de recuperacion de dinucleotido de nicotinamida adenina cuaternizados.
BR112019023014A2 (pt) 2017-05-02 2020-05-19 Immunomic Therapeutics Inc constructos de lamp aperfeiçoados compreendendo antígenos de câncer
CN111010867B (zh) 2017-05-30 2023-05-12 俄克拉何马大学董事会 抗双皮层蛋白样激酶1抗体及其使用方法
EP3431496A1 (de) 2017-07-19 2019-01-23 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Anti-isoasp7-amyloid-beta-antikörper und verwendungen davon
WO2019020480A1 (en) 2017-07-24 2019-01-31 INSERM (Institut National de la Santé et de la Recherche Médicale) ANTIBODIES AND PEPTIDES FOR TREATING HCMV RELATED DISEASES
BR112020005671A2 (pt) 2017-09-21 2020-10-20 INSERM (Institut National de la Santé et de la Recherche Médicale) anticorpos que apresentam especificidade para btn2 e usos dos mesmos
WO2019067015A1 (en) 2017-09-29 2019-04-04 City Of Hope RECEPTORS OF CHIMERIC ANTIGENS AND BISPECIFIC ANTIBODIES FOR THE TREATMENT OF COAT CELL LYMPHOMA
BR112020005419A2 (pt) 2017-10-02 2020-09-29 Visterra, Inc. moléculas de anticorpo para cd138 e seus usos
US10640508B2 (en) 2017-10-13 2020-05-05 Massachusetts Institute Of Technology Diazene directed modular synthesis of compounds with quaternary carbon centers
WO2019077164A1 (en) 2017-10-20 2019-04-25 Institut Curie CROCHET FUSION PROTEIN FOR REGULATING THE CELLULAR TRAFFIC OF A TARGET PROTEIN
US20200331975A1 (en) 2017-10-20 2020-10-22 Institut Curie Dap10/12 based cars adapted for rush
IL276546B2 (en) 2018-02-20 2024-08-01 Seagen Inc Hydrophobic auristatin F compounds and their conjugates
EP3530282A1 (de) 2018-02-27 2019-08-28 Diaccurate Therapeutische verfahren
CA3100004A1 (en) 2018-05-15 2019-11-21 Immunomic Therapeutics, Inc. Improved lamp constructs comprising allergens
TWI851577B (zh) 2018-06-07 2024-08-11 美商思進公司 喜樹鹼結合物
EP3807316B1 (de) 2018-06-18 2024-05-01 Innate Pharma Zusammensetzungen und verfahren zur behandlung von krebs
SG11202012712YA (en) 2018-06-19 2021-01-28 Atarga Llc Antibody molecules to complement component 5 and uses thereof
WO2020006568A1 (en) 2018-06-29 2020-01-02 City Of Hope Cd6 targeted chimeric antigen receptors for treatent of certain autoimmune disorders
WO2020010250A2 (en) 2018-07-03 2020-01-09 Elstar Therapeutics, Inc. Anti-tcr antibody molecules and uses thereof
CN112867503A (zh) 2018-08-24 2021-05-28 希望之城 掩蔽的细胞因子缀合物
WO2020053122A1 (en) 2018-09-10 2020-03-19 INSERM (Institut National de la Santé et de la Recherche Médicale) Combination of her2/neu antibody with heme for treating cancer
WO2020053808A1 (en) 2018-09-12 2020-03-19 Georg Dewald Method of diagnosing vasoregulatory disorders
EP3853250A4 (de) 2018-09-19 2022-06-08 La Jolla Institute for Immunology Ptprs und proteoglykane in rheumatoider arthritis
EP3860641A1 (de) 2018-10-05 2021-08-11 Bavarian Nordic A/S Kombinationstherapie zur behandlung von krebs mit einer intravenösen verabreichung eines rekombinanten mva und eines immunkontrollpunktantagonisten oder -agonisten
AU2019385665A1 (en) 2018-11-20 2021-05-27 Bavarian Nordic A/S Therapy for treating cancer with an intratumoral and/or intravenous administration of a recombinant MVA encoding 4-1BBL (CD137L) and/or CD40l
EP3693063A1 (de) 2019-02-06 2020-08-12 Diaccurate Verfahren und zusammensetzungen zur behandlung von krebs
EP3924379A4 (de) 2019-02-15 2022-12-21 Integral Molecular, Inc. Antikörper mit einer gemeinsamen leichten kette und deren verwendungen
EP3924389A4 (de) 2019-02-15 2023-06-14 Integral Molecular, Inc. Claudin-6-antikörper und deren verwendungen
US12109273B2 (en) 2019-02-15 2024-10-08 Wuxi Xdc Singapore Private Limited Process for preparing antibody-drug conjugates with improved homogeneity
KR20210125034A (ko) 2019-02-15 2021-10-15 우시 바이올로직스 아일랜드 리미티드 동질성이 개선된 항체-약물 콘쥬게이트의 제조방법
CA3134122A1 (en) 2019-03-20 2020-09-24 Imcheck Therapeutics Sas Antibodies having specificity for btn2 and uses thereof
CN113631573B (zh) 2019-03-25 2024-06-04 国家医疗保健研究所 抗Tau抗体及其在制备用于治疗疾病的药物中的用途
US20220185876A1 (en) 2019-03-29 2022-06-16 Atarga, Llc Anti fgf23 antibody
BR112021022514A2 (pt) 2019-05-10 2022-04-19 Takeda Pharmaceuticals Co Conjugados de anticorpo e fármaco
US20200392241A1 (en) 2019-06-17 2020-12-17 Visterra, Inc. Humanized antibody molecules to cd138 and uses thereof
WO2021023624A1 (en) 2019-08-02 2021-02-11 Orega Biotech Novel il-17b antibodies
AU2020348876A1 (en) 2019-09-19 2022-04-07 Seagen Inc. Selective drug release from internalized conjugates of biologically active compounds
JP2022550325A (ja) 2019-09-27 2022-12-01 アンスティチュ ナショナル ドゥ ラ サンテ エ ドゥ ラ ルシェルシュ メディカル 抗ミュラー管抑制物質抗体およびその使用
WO2021058729A1 (en) 2019-09-27 2021-04-01 INSERM (Institut National de la Santé et de la Recherche Médicale) Anti-müllerian inhibiting substance type i receptor antibodies and uses thereof
JP2022550851A (ja) 2019-10-04 2022-12-05 シージェン インコーポレイテッド カンプトテシンペプチドコンジュゲート
WO2021067820A1 (en) 2019-10-04 2021-04-08 Seagen Inc. Formulation of antibody-drug conjugate
KR20220083773A (ko) 2019-10-18 2022-06-20 이뮤노믹 쎄라퓨틱스, 인크. 암 항원을 포함하는 개선된 lamp 구축물
MX2022005664A (es) 2019-11-20 2022-09-07 Bavarian Nordic As Virus de vaccinia ankara modificados (mva) recombinantes para administración intratumoral y/o intravenosa para tratamiento de cáncer.
EP4072682A1 (de) 2019-12-09 2022-10-19 Institut National de la Santé et de la Recherche Médicale (INSERM) Antikörper mit spezifität gegen her4 und verwendungen davon
EP4084808A1 (de) 2019-12-30 2022-11-09 City of Hope Verfahren zur herstellung und verwendung von regulatorischen t-zellen und effektor-t-zellen mit gegen cd6, cd19, und/oder il-13r gerichteten chimären antigen-rezeptoren zur behandlung von autoimmunerkrankungen und krebs
TW202138388A (zh) 2019-12-30 2021-10-16 美商西根公司 以非海藻糖苷化抗－ｃｄ７０抗體治療癌症之方法
TW202140553A (zh) 2020-01-13 2021-11-01 美商威特拉公司 C5ar1抗體分子及其用途
US12029736B2 (en) 2020-02-25 2024-07-09 Mediboston Limited Camptothecin derivatives and conjugates thereof
WO2021175954A1 (en) 2020-03-04 2021-09-10 Imcheck Therapeutics Sas Antibodies having specificity for btnl8 and uses thereof
US20210311054A1 (en) 2020-04-01 2021-10-07 Institut Pasteur Severe acute respiratory syndrome (sars) - associated coronavirus diagnostics
US11815513B2 (en) 2020-04-01 2023-11-14 Institut Pasteur Severe acute respiratory syndrome (SARS)-associated coronavirus diagnostics
MX2022012621A (es) 2020-04-10 2023-01-18 Seagen Inc Enlazadores de variantes de carga.
WO2021209824A1 (en) 2020-04-17 2021-10-21 Institut Pasteur Methods and products for serological analysis of sars-cov-2 infection
KR20230005258A (ko) 2020-04-24 2023-01-09 사노피 항-ceacam5 항체 접합체 및 트리플루리딘 및 티피라실을 함유하는 항종양 병용물
CN115768484A (zh) 2020-04-24 2023-03-07 赛诺菲 含有抗ceacam5抗体缀合物和伊立替康的抗肿瘤组合
WO2021214227A1 (en) 2020-04-24 2021-10-28 Sanofi Antitumor combinations containing anti-ceacam5 antibody conjugates and cetuximab
MX2022013405A (es) 2020-04-24 2022-11-14 Sanofi Sa Combinaciones antitumorales que contienen productos conjugados de anticuerpos anti-ceacam5 y folfox.
WO2021220218A1 (en) 2020-05-01 2021-11-04 Novartis Ag Immunoglobulin variants
JP2023523794A (ja) 2020-05-01 2023-06-07 ノバルティス アーゲー 人工操作免疫グロブリン
KR20230008751A (ko) 2020-05-12 2023-01-16 인쎄름 (엥스띠뛰 나씨오날 드 라 쌍떼 에 드 라 흐쉐르슈 메디깔) 피부 t-세포 림프종 및 tfh 유래된 림프종을 치료하는 신규한 방법
WO2021239666A1 (en) 2020-05-26 2021-12-02 Diaccurate Therapeutic methods
EP4172206A1 (de) 2020-06-24 2023-05-03 Visterra, Inc. Antikörpermoleküle gegen april und verwendungen davon
EP4178678A1 (de) 2020-07-07 2023-05-17 Cancure, LLC Mic-antikörper und bindemittel und verfahren zur verwendung davon
CA3184852A1 (en) 2020-07-10 2022-01-13 Lida Katsimpardi Use of gdf11 to diagnose and treat anxiety and depression
WO2022026592A2 (en) 2020-07-28 2022-02-03 Celltas Bio, Inc. Antibody molecules to coronavirus and uses thereof
WO2022035998A1 (en) 2020-08-11 2022-02-17 City Of Hope Compositions and uses of sars-cov-2 targeted chimeric antigen receptor modified nk cells
MX2023002330A (es) 2020-09-04 2023-03-21 Merck Patent Gmbh Anticuerpos anti molecula de adhesion celular 5 relacionada con el antigeno carcinoembrionario (ceacam5) y conjugados y usos de los mismos.
EP4208197A1 (de) 2020-09-04 2023-07-12 Rutgers, The State University of New Jersey Sars-cov-2-impfstoffe und -antikörper
EP4214223A1 (de) 2020-09-21 2023-07-26 Theravectys Verfahren und produkte mit hohem durchsatz für sars-cov-2-seroneutralisierungsassay
EP4229081A1 (de) 2020-10-15 2023-08-23 The United States of America, as represented by The Secretary, Department of Health and Human Services Für sars-cov-2-rezeptor-bindungsdomäne spezifischer antikörper und therapeutische verfahren
WO2022087274A1 (en) 2020-10-21 2022-04-28 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Antibodies that neutralize type-i interferon (ifn) activity
AU2021374958A1 (en) 2020-11-09 2023-06-22 Takeda Pharmaceutical Company Limited Antibody drug conjugates
AU2022208054A1 (en) 2021-01-15 2023-07-27 Seagen Inc. Immunomodulatory antibody-drug conjugates
CA3208974A1 (en) 2021-01-20 2022-07-28 Bioentre Llc Ctla4-binding proteins and methods of treating cancer
WO2022169825A1 (en) 2021-02-03 2022-08-11 Mozart Therapeutics, Inc. Binding agents and methods of using the same
CN116847886A (zh) 2021-02-03 2023-10-03 思进公司 免疫刺激化合物及缀合物
TW202300178A (zh) 2021-03-18 2023-01-01 美商西根公司 內化的生物活性化合物偶聯物選擇性釋放藥物
AU2022237791A1 (en) 2021-03-18 2023-09-14 Seagen Inc. Selective drug release from internalized conjugates of biologically active compounds
TW202304524A (zh) 2021-04-10 2023-02-01 美商普方生物製藥美國公司 Folr1結合劑、其結合物及使用方法
WO2022226317A1 (en) 2021-04-23 2022-10-27 Profoundbio Us Co. Anti-cd70 antibodies, conjugates thereof and methods of using the same
KR20240015670A (ko) 2021-05-28 2024-02-05 씨젠 인크. 안트라사이클린 항체 접합체
TW202317190A (zh) 2021-06-29 2023-05-01 美商思進公司 以非岩藻糖基化抗ｃｄ７０抗體及ｃｄ４７拮抗劑之組合治療癌症之方法
TW202320857A (zh) 2021-07-06 2023-06-01 美商普方生物製藥美國公司 連接子、藥物連接子及其結合物及其使用方法
KR20240095160A (ko) 2021-07-29 2024-06-25 인스티튜트 내셔널 드 라 싼테 에 드 라 리셰르셰 메디칼르 (인 썸) 인간화된 항-인간 βig-h3 단백질 및 이의 용도
WO2023076876A1 (en) 2021-10-26 2023-05-04 Mozart Therapeutics, Inc. Modulation of immune responses to viral vectors
WO2023079057A1 (en) 2021-11-05 2023-05-11 Sanofi Antitumor combinations containing anti-ceacam5 antibody-drug conjugates and anti-vegfr-2 antibodies
CA3238167A1 (en) 2021-11-19 2023-05-25 Maria Leia Smith Gpc3 binding agents, conjugates thereof and methods of using the same
WO2023097254A1 (en) 2021-11-24 2023-06-01 Visterra, Inc. Engineered antibody molecules to cd138 and uses thereof
EP4452305A1 (de) 2021-12-23 2024-10-30 Bavarian Nordic A/S Rekombinante mva-viren zur intraperitonealen verabreichung zur behandlung von krebs
WO2023150181A1 (en) 2022-02-01 2023-08-10 President And Fellows Of Harvard College Methods and compositions for treating cancer
US20230383010A1 (en) 2022-02-07 2023-11-30 Visterra, Inc. Anti-idiotype antibody molecules and uses thereof
WO2023172968A1 (en) 2022-03-09 2023-09-14 Merck Patent Gmbh Anti-gd2 antibodies, immunoconjugates and therapeutic uses thereof
WO2023170239A1 (en) 2022-03-09 2023-09-14 Merck Patent Gmbh Methods and tools for conjugation to antibodies
IL315508A (en) 2022-03-17 2024-11-01 Seagen Inc Camptothecin conjugates
WO2023201201A1 (en) 2022-04-10 2023-10-19 Immunomic Therapeutics, Inc. Bicistronic lamp constructs comprising immune response enhancing genes and methods of use thereof
WO2023212518A1 (en) 2022-04-25 2023-11-02 Visterra, Inc. Antibody molecules to april and uses thereof
WO2023215740A1 (en) 2022-05-06 2023-11-09 Seagen Inc. Immunomodulatory antibody-drug conjugates
WO2023223092A1 (en) 2022-05-18 2023-11-23 Institut Pasteur Identification of a human circovirus
WO2023240287A1 (en) 2022-06-10 2023-12-14 Bioentre Llc Combinations of ctla4 binding proteins and methods of treating cancer
WO2024023735A1 (en) 2022-07-27 2024-02-01 Mediboston Limited Auristatin derivatives and conjugates thereof
WO2024030577A1 (en) 2022-08-03 2024-02-08 Seagen Inc. Immunostimulatory anti-pd-l1-drug conjugates
WO2024030956A2 (en) 2022-08-03 2024-02-08 Mozart Therapeutics, Inc. Cd39-specific binding agents and methods of using the same
EP4321522A1 (de) 2022-08-12 2024-02-14 Seagen Inc. Zytotoxische verbindungen und konjugate davon
WO2024052503A1 (en) 2022-09-08 2024-03-14 Institut National de la Santé et de la Recherche Médicale Antibodies having specificity to ltbp2 and uses thereof
WO2024056668A1 (en) 2022-09-12 2024-03-21 Institut National de la Santé et de la Recherche Médicale New anti-itgb8 antibodies and its uses thereof
TW202432099A (zh) 2022-10-25 2024-08-16 美商默沙東有限責任公司 源自依克沙替康(exatecan)之adc連接子-載藥(payload)、醫藥組合物及其用途
WO2024118771A1 (en) 2022-11-30 2024-06-06 Integral Molecular, Inc. Antibodies directed to claudin 6, including bispecific formats thereof
WO2024129756A1 (en) 2022-12-13 2024-06-20 Seagen Inc. Site-specific engineered cysteine antibody drug conjugates
WO2024129628A1 (en) 2022-12-14 2024-06-20 Merck Sharp & Dohme Llc Auristatin linker-payloads, pharmaceutical compositions, and uses thereof
WO2024168061A2 (en) 2023-02-07 2024-08-15 Ayan Therapeutics Inc. Antibody molecules binding to sars-cov-2
WO2024180192A1 (en) 2023-03-01 2024-09-06 Sanofi Use of anti-ceacam5 immunoconjugates for treating neuroendocrine cancers expressing ceacam5
EP4427763A1 (de) 2023-03-06 2024-09-11 Sanofi Antitumorkombinationen mit anti-ceacam5-antikörper-wirkstoff-konjugaten, anti-vegfr-2-antikörpern und anti-pd1/pd-l1-antikörpern
WO2024194455A1 (en) 2023-03-23 2024-09-26 Sanofi CEACAM5 mRNA ASSAY FOR PATIENT SELECTION IN CANCER THERAPY

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8308235D0 (en) * 1983-03-25 1983-05-05 Celltech Ltd Polypeptides
US4816567A (en) * 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
JPS6147500A (ja) * 1984-08-15 1986-03-07 Res Dev Corp Of Japan キメラモノクロ−ナル抗体及びその製造法
EP0173494A3 (de) * 1984-08-27 1987-11-25 The Board Of Trustees Of The Leland Stanford Junior University Chimäre Rezeptoren durch Verbindung und Expression von DNS
GB8422238D0 (en) * 1984-09-03 1984-10-10 Neuberger M S Chimeric proteins
JPS61134325A (ja) * 1984-12-04 1986-06-21 Teijin Ltd ハイブリツド抗体遺伝子の発現方法
JPS62502586A (ja) * 1985-03-18 1987-10-08 ジ−ン　ラブズ，インコ−ポレイテツド ハイブリツド遺伝子カセツトベクタ−

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
NATURE (London) Vol. 314 Issued 21 March 1985 (NEUBERGER ET AL) "A Hapten-Specific Chimaeric IgE Antibody with Human Physiological Effector Function" s. pp. 268-270. *
NATURE (London) Vol. 314 Issued 4 April 1985 (TAKEDA ET AL) "Construction of Chimaeric Processed Immunoglobulin Genes Containing Mouse Variable and Human Constant Region Sequences", s. pp. 452-454 *

Also Published As

Publication number Publication date
AU606320B2 (en) 1991-02-07
AU6598186A (en) 1987-05-19
DE3689123D1 (de) 1993-11-04
EP0247091A1 (de) 1987-12-02
WO1987002671A1 (en) 1987-05-07
JPS63501765A (ja) 1988-07-21
DK338587D0 (da) 1987-07-01
DK338587A (da) 1987-07-01
EP0247091A4 (de) 1987-10-27
DE3689123T2 (de) 1994-03-03
DK175680B1 (da) 2005-01-17

Similar Documents

Publication Publication Date Title
EP0247091B1 (de) Modulare einheit von antikörpergenen, daraus hergestellte antikörper und verwendung
EP0371998B1 (de) Modulärer zusammenbau von antikörpergenen, dadurch hergestellte antikörper und deren anwendung
US5500362A (en) Chimeric antibody with specificity to human B cell surface antigen
EP0266663B1 (de) Chimärer Antikörper mit Spezifität für menschliches Tumor-Antigen
US5595898A (en) Modular assembly of antibody genes, antibodies prepared thereby and use
US5846818A (en) Pectate lyase signal sequence
US6893625B1 (en) Chimeric antibody with specificity to human B cell surface antigen
EP0536566A1 (de) Modulare Einheit von Antikörpergenen, daraus hergestellte Antikörper und Verwendung
DK175581B1 (da) Modulær samling af antistofgener
AU606653C (en) Chimeric antibody with specificity to human B cell surface antigen

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

A4 Supplementary search report drawn up and despatched

Effective date: 19871027

17P Request for examination filed

Effective date: 19871106

17Q First examination report despatched

Effective date: 19881027

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: XOMA CORPORATION

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH DE FR GB IT LI LU NL SE

REF Corresponds to:

Ref document number: 95235

Country of ref document: AT

Date of ref document: 19931015

Kind code of ref document: T

XX Miscellaneous (additional remarks)

Free format text: TEILANMELDUNG 92115754.1 EINGEREICHT AM 27/10/86.

ET Fr: translation filed
REF Corresponds to:

Ref document number: 3689123

Country of ref document: DE

Date of ref document: 19931104

ITF It: translation for a ep patent filed
EPTA Lu: last paid annual fee
PLBI Opposition filed

Free format text: ORIGINAL CODE: 0009260

26 Opposition filed

Opponent name: GENENTECH, INC.

Effective date: 19940627

Opponent name: CELLTECH LIMITED

Effective date: 19940623

NLR1 Nl: opposition has been filed with the epo

Opponent name: GENETECH, INC.

Opponent name: CELLTECH LIMITED.

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 19941130

Year of fee payment: 9

EAL Se: european patent in force in sweden

Ref document number: 86906676.1

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: LU

Payment date: 19951001

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 19951006

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 19951010

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 19951013

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: SE

Payment date: 19951017

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 19951026

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: AT

Payment date: 19951027

Year of fee payment: 10

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 19951031

Year of fee payment: 10

RDAG Patent revoked

Free format text: ORIGINAL CODE: 0009271

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: PATENT REVOKED

27W Patent revoked

Effective date: 19951121

GBPR Gb: patent revoked under art. 102 of the ep convention designating the uk as contracting state

Free format text: 951121

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

NLR2 Nl: decision of opposition
PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20051116

Year of fee payment: 20